Clk-2, cex-7 and coq-4 genes, and uses thereof

ABSTRACT

The present invention relates to a clk-2 gene which has a function at the level of cellular physiology involved in developmental rate, telomere length and longevity, wherein clk-2 mutations cause a longer life, an altered cellular metabolism and an altered telomere length relative to the wild type, wherein clk-2 overexpression leads to telomere shortening. The present invention also relates to clk-2 co-expressed gene which comprises a cex-7 gene having the nucleotide sequence set forth in FIG.  33  which codes for a CEX-7 protein having the amino acid sequence set forth in FIG.  34  wherein said gene is located in the clk-2 operon and said cex-7 gene is transcriptionally co-expressed with clk-2 gene present in said operon. The present invention also relates to a coq-4 gene which has a function at the level of cellular physiology involved in the regulation of developmental rate and longevity, wherein coq-4 mutations cause altered cellular metabolism and physiological relative to the wild type, wherein coq-4 gene has the identifying characteristics of nucleotide sequence set forth in FIG.  36.

BACKGROUND OF THE INVENTION

[0001] (a) Field of the Invention

[0002] The invention relates to the identification of three genes: the gene clk-2, the gene cex-7 that is located in the same operon as clk-2, and the gene coq-4. The invention shows that these genes regulate the timing of development and behavior, and determine life span and that clk-2 regulates the length of telomeres.

[0003] (b) Description of Prior Art

[0004] A class of genes was identified in the nematode Caenorhabditis elegans, the clk (‘clock’) genes, whose activity controls how fast the worms live and die. Mutations in these genes result in an alteration of developmental and behavioral timing, including an average slow down of the animal's embryonic and post-embryonic development and of their rhythmic behaviors, as well as an increase in the animal's life span. In addition, mutations in these genes display a maternal effect, namely, homozygous mutants (clk/clk) derived from a heterozygous mother (clk/+), appear phenotypically wild-type.

[0005] We isolated the mutations that define the genes clk-1, clk-2, clk-3 in a screen for viable maternal-effect mutations in the nematode Caenorhabditis elegans (Hekimi, S. et al., Genetics 141, 1351 (1995)). gro-1 was originally identified by a spontaneous mutation isolated from a strain that had been recently established from a wild isolate (Hodgkin, J. and Doniach, T. Genetics 146, 149 (1997)). Our subsequent reappraisal of this mutation revealed that it shares the characteristics of the clk genes (Wong, A. et al., Genetics 139, 1247 (1995)).

[0006] We have molecularly identified two of these genes, clk-1 and gro-1. clk-1 encodes a protein that is highly conserved from proteobacteria to humans which is structurally similar to the yeast metabolic regulator Cat5p/Coq7p (Ewbank, J. J. et al, Science 275, 980 (1997); WO98/17823). gro-1 encodes a highly conserved cellular enzyme, the dimethylallyltransferase:tRNA dimethylallyltransferase (WO99/10482).

[0007] To date, clk-1 is the gene that has been characterized in greatest detail. In addition to the phenotypic and molecular characterization, it was found that clk-1 is ubiquitously expressed in the worm's body where it localizes to the mitochondria, the energy generating organelle of the cell (Felkai, S. et al, EMBO Journal 18, 1783 (1999)). clk-1 thus controls timing by regulating physiological rates (Branicky R, C. Benard, S. Hekimi, Bioessays 22, 48 (2000)).

[0008] The gene clk-2 is defined by one allele that was isolated in a screen for viable maternal-effect mutations in Caenorhabditis elegans (Hekimi, S. et al., Genetics 141, 1351 (1995)). The mutations in the gene clk-2 were shown to result in an alteration of the timing of several developmental and behavioral events (Hekimi, S. et al., Genetics 141, 1351 (1995)) and that the activity of the gene clk-2 controls how fast the worms live and how soon they die (Lakowski, B. and Hekimi, S. Science 272, 1010 (1996). We have also noticed other phenotypes of the clk-2 mutants such as the temperature sensitivity of the clk-2 (qm37) allele. Overall, these phenotypes are similar to those of mutations in the three clk genes (Hekimi, S. et al., Genetics 141, 1351 (1995)).

[0009] Results to date suggest that the effect of clk genes on the rate of aging is due to an effect on the rate of living. First, clk-1 mutations which lead to a decrease of clk-1 activity result in a slow down of development and behavior and in an increase in life span. On the other hand, overexpression of clk-1 in transgenic animals accelerates the rate of living as revealed by the absence of a characteristic behavioral slow down with age. Second, the effect of the different clk genes is additive. We have shown that double clk mutants develop more slowly and live longer than the single clk mutants. Third, clk genes are distinct from dauer formation genes (daf genes) which are involved in stress resistance and also prolong life span. Daf genes affect life span through a separate mechanism from that of clk. In fact, clk mutants are neither dauer constitutive nor dauer defective and daf-16 mutations cannot suppress the long life of clk-1, -2, -3 mutants.

[0010] The gene coq-4 is similar to the gene clk-1 in that both genes are required for normal ubiquinone biosynthesis in yeast and both genes have no homologues in E. coli. The gene cex-7 that will be described below has been found to be a pseudoautosomal gene named XE7 in humans.

[0011] It would be highly desirable to be provided with a detailed phenotypic and molecular characterization of the gene clk-2, as well as with a characterization of the gene coq-4 in an animal.

SUMMARY OF THE INVENTION

[0012] One aim of the present invention is to provide with a clk-2 gene which has a function at the level of cellular physiology involved in developmental rate, telomere length and longevity, wherein clk-2 mutations cause a longer life, an altered cellular metabolism and an altered telomere length relative to the wild type, wherein clk-2 gene has the identifying characteristics of nucleotide sequence described in FIG. 1.

[0013] In accordance with the present invention there is provided the use of a clk-2 gene to alter a function at the level of cellular physiology involved in the regulation of developmental rates, telomere length and longevity, wherein clk-2 mutations cause a longer life, an altered cellular metabolism and physiological rates and an altered telomere length relative to the wild type, wherein clk-2 gene has the identifying characteristics of nucleotide sequences described in FIGS. 1, 4-7, 15, 16, and 20-24, or wherein the gene codes for a protein sequence as described in FIGS. 2, 3, 8-14, 17-19, and 25-32 as deduced from FIGS. 1, 4-7, 15, 16, 20-24.

[0014] Also is provided with the invention the use of a clk-2 gene to alter function at the level of cellular physiology involved in the regulation of developmental rate, telomere length and longevity, wherein clk-2 mutations cause a longer life and altered cellular metabolism and physiological rates and an altered telomere length relative to the wild type, wherein the gene codes for a protein having a sequence as set forth in FIG. 32.

[0015] In accordance with the invention there is provided a CLK-2 protein that has a function at the level of cellular physiology involved in the regulation of developmental rate, telomere length and longevity.

[0016] There is also provided with the invention a mutant CLK-2 protein which has the amino acid sequence described in FIG. 31, and the use of CLK-2 protein to alter a function at the level of cellular physiology involved in the regulation of developmental rates, telomere length and longevity, wherein the CLK-2 protein has the amino acid sequence as described in FIGS. 2, 3, 8-14, 17-19, and 25-32.

[0017] In accordance with the invention, there is provided a clk-2 gene which has the nucleotide sequence described in FIG. 1, and the use of clk-2 gene and homologues thereof, to manipulate the physiological rates and/or telomere biology, whereby life span of an organism is altered.

[0018] There is also provided a mouse which comprises a gene knockout of the murine clk-2 gene homologue to a clk-2 gene.

[0019] There is provided with the present invention a method to increase the life span of multicellular organism and metazoan which comprises altering the function of telomeres and mechanisms of sub-telomeric silencing.

[0020] The invention also provides the use of clk-2 gene, CLK-2 protein, and homologues thereof, for screening drugs which decrease or increase the life span of a multicellular organism, wherein the drug enhances or suppresses the expression of the clk-2 gene or activity of the protein CLK-2, and homologues thereof.

[0021] The use of a compound is provided with the invention for the manufacture of a medicament for increasing and/or decreasing physiological rates of tissues, organ, and/or whole organism of a host; wherein the compound is interfering with activity of CLK-2 protein and homologues thereof.

[0022] The use of a compound is also provided to promote tissue and/or organ specific reduction or increase of clk-2 activity for the manufacture of a medicament for the treatment of pathological conditions causing increase or decrease of physiological rate of tissue and/or organ in an individual, wherein the compound is interfering with activity of CLK-2 protein and homologues thereof.

[0023] In accordance with the invention there is provided a clk-2 co-expressed gene which comprises a cex-7 gene having the nucleotide sequence as described in FIG. 33, and which codes for a CEX-7 protein having the amino acid sequence described in FIG. 34 wherein the gene is located in the clk-2 operon and the cex-7 gene is transcriptionally co-expressed with clk-2 gene present in the same operon.

[0024] A human homologue of cex-7 gene is also provided with the invention, wherein the gene codes for a protein having a sequence as described in FIG. 35.

[0025] There is provided with the invention the use of a human homologue of cex-7 gene and homologues thereof to alter a function at the level of cellular level physiology involved in the regulation of developmental rates and longevity wherein the gene codes for a protein having a sequence as described in FIG. 35.

[0026] The invention provides with a mouse which comprises a gene knock out of the murine cex-7 gene homologue of the human gene described in FIG. 35.

[0027] There is also provided with the invention the use of a compound for the manufacture of a medicament for increasing and/or decreasing physiological rates of tissues, organ, and/or whole organism of a host; wherein the compound is interfering with activity of CEX-7 and homologues thereof.

[0028] Another aim of the invention is to provide with the use of a compound which promotes tissue and/or organ specific reduction or increase of cex-7 activity for the manufacture of a medicament for the treatment of pathological conditions causing increase or decrease of physiological rate of tissue and/or organ in an individual, wherein the compound is interfering with activity of CEX-7 and homologues thereof.

[0029] There is provided with the invention a coq-4 gene which has a function at the level of cellular physiology involved in the regulation of developmental rate and longevity, wherein coq-4 mutations cause altered cellular metabolism and physiological relative to the wild type, wherein coq-4 gene has the identifying characteristics of nucleotide sequence as described in FIG. 36.

[0030] A coq-4 gene provided with the invention has a function at the level of cellular physiology involved in the regulation of developmental rate and longevity, wherein coq-4 mutations cause altered cellular metabolism and physiological relative to the wild type, wherein coq-4 gene has the identifying characteristics of nucleotide sequence as described in FIG. 36, and the gene codes for a protein having a sequence as described in FIG. 37.

[0031] In accordance with the invention, there is provided with the use of coq-4 gene to alter a function at the level of cellular physiology involved in the regulation of developmental rates, wherein coq-4 mutations cause an altered cellular metabolism and physiological rates relative to the wild type, wherein the gene codes for a protein having a sequence as described in FIGS. 43 to 54 and homologues thereof.

[0032] There is also provided with the invention a mouse which comprises a gene knock out of the murine coq-4 gene as described in FIG. 47.

[0033] There is provided also with the invention the use of a compound for the manufacture of a medicament for increasing and/or decreasing physiological rates of tissues, organs and/or whole organism of a host; wherein the compound is interfering with activity of COQ-4 and homologues thereof.

[0034] A compound in accordance with the invention is provided to promote tissue and/or organ specific reduction or increase of coq-4 activity for the manufacture of a medicament for the treatment of pathological conditions causing increase or decrease of physiological rate of tissue and/or organ in an individual, wherein the compound is interfering with activity of COQ-4 and homologues thereof.

[0035] Having the clk genes in hand can serve to manipulate the rate of development, the cell cycle, the rate of behavior and the rate of aging. Another way to look at it is that it can help to control physiological rates including for medical and industrial purposes. Slowing down the rate of aging of individual organs or tissues to slow down their rate of deterioration is one medical example; accelerating the growth of farm animals or crops is an example of industrial utilization.

[0036] Here we describe our analysis of clk-2 and cex-7 and the inventions that result from this analysis, including the molecular characterization of clk-2 and cex-7 and the identification of homologues in several species, including humans. We also describe our identification of a new clk gene: coq-4. We have obtained a mutation in the worm coq-4 locus and have shown that the mutant animals display several of the most important characteristics of clk mutations.

BRIEF DESCRIPTION OF THE DRAWINGS

[0037]FIGS. 1A and 1B illustrate the Caenorhabditis elegans clk-2 cDNA sequence.

[0038]FIG. 2 illustrates the Caenorhabditis elegans CLK-2 protein sequence.

[0039]FIG. 3 illustrates the Homo sapiens CLK-2 protein sequence(derived from clone KIAA0683).

[0040]FIGS. 4A and 4B illustrate the Homo sapiens clk-2 homologue nucleotide sequence (derived from AL080126).

[0041]FIG. 5 illustrates part of Mus musculus clk-2 cDNA sequence (derived from AA671905 vl11b10.r1).

[0042]FIG. 6 illustrates part of Mus musculus clk-2 cDNA sequence (derived from AA031108 mi40f03.r1).

[0043]FIG. 7 illustrates part of Mus musculus clk-2 cDNA sequence (derived from AA230994 mw30h11.r1).

[0044]FIG. 8 illustrates part of Mus musculus CLK-2 protein sequence (derived from gb|AA671905.1|AA671905.)

[0045]FIG. 9 illustrates part of Mus musculus CLK-2 protein sequence (derived from gb|AA230994.1|AA230994).

[0046]FIG. 10 illustrates part of Mus musculus CLK-2 protein sequence (derived from gb|AA031108.1|AA031108).

[0047]FIG. 11 illustrates Mus musculus composite CLK-2 protein sequence.

[0048]FIG. 12 illustrates part of Sus scrofa CLK-2 protein sequence (derived from gb|AW429611.1|AW429611).

[0049]FIG. 13 illustrates the Drosophila melanogaster CLK-2 protein sequence.

[0050]FIG. 14 illustrates the putative Arabidopsis thaliana CLK-2 protein sequence (derived from 7630034|emb|CAB88328.1|).

[0051]FIG. 15 illustrates part of Oryza sativa clk-2 cDNA sequence (derived from AU031811).

[0052]FIG. 16 illustrates part of Oryza sativa clk-2 cDNA sequence (derived from D24238).

[0053]FIG. 17 illustrates part of Oryza sativa CLK-2 protein sequence (derived from dbj|D24422.1|D24422).

[0054]FIG. 18 illustrates part of Oryza sativa CLK-2 protein sequence (derived from dbj|AU031811.1|AU031811).

[0055]FIG. 19 illustrates Oryza sativa composite CLK-2 protein.

[0056]FIG. 20 illustrates part of Glycine max clk-2 cDNA sequence (derived from AI461201 sa76d07.y1 Gm-c1004).

[0057]FIG. 21 illustrates part of Glycine max clk-2 cDNA sequence (derived from AW185029 se85g06.y1 Gm-c1023).

[0058]FIG. 22 illustrates part of Glycine max clk-2 cDNA sequence (derived from AW350166 GM210007A10F4R Gm-r1021).

[0059]FIG. 23 illustrates part of Glycine max clk-2 cDNA sequence (derived from AW397826 sg68g12.y1 Gm-c1007).

[0060]FIG. 24 illustrates part of Glycine max clk-2 cDNA sequence (derived from AW567713 si54a01.y1 Gm-r1030).

[0061]FIG. 25 illustrates part of Glycine max CLK-2 protein sequence (derived from gb|AW350166.1|AW350166).

[0062]FIG. 26 illustrates part of Glycine max CLK-2 protein sequence (derived from gb|AI461201.1|AI461201).

[0063]FIG. 27 illustrates part of Glycine max CLK-2 protein sequence (derived from gb|AW|85029.1|AW185029).

[0064]FIG. 28 illustrates part of Glycine max CLK-2 protein sequence (derived from gb|AW567713.1|AW567713).

[0065]FIG. 29 illustrates part of Glycine max CLK-2 protein sequence (derived from gb|AW397826.1|AW397826).

[0066]FIG. 30 illustrates Glycine max CLK-2 composite protein sequence.

[0067]FIG. 31 illustrates the Caenorhabditis elegans CLK-2 (QM37) mutant protein, with C to Y substitution at position 772.

[0068]FIG. 32 illustrates Tel2p, the Saccharomyces cerevisiae CLK-2 protein.

[0069]FIG. 33 illustrates the Caenorhabditis elegans cex-7 cDNA sequence.

[0070]FIG. 34 illustrates the Caenorhabditis elegans CEX-7 protein sequence.

[0071]FIG. 35 illustrates the Homo sapiens CEX-7 protein sequence (XE7).

[0072]FIG. 36 illustrates the Caenorhabditis elegans coq-4 cDNA sequence.

[0073]FIG. 37 illustrates the Caenorhabditis elegans COQ-4 protein sequence.

[0074]FIGS. 38A and 38B illustrate the comparison of CLK-2 eukaryotic homologues (hCLK-2: Homo sapiens CLK-2: Caenorhabditis elegans Tel2p: Saccharomyces cerevisiae AtCLK-2: Arabidopsis thaliana).

[0075]FIG. 39 illustrates the comparison of CLK-2 animal homologues (D.m.: Drosophila melanogaster, H.s.: Homo sapiens C.e.: Caenorhabditis elegans).

[0076]FIG. 40 illustrates the comparison of CLK-2 vertebrate homologues (H.s.: Homo sapiens, M.m.: Mus musculus, S.s.: Sus scrofa).

[0077]FIG. 41 illustrates the comparison of CLK-2 plant homologues (A.t.: Arabidopsis thaliana, G.m.: Glycine max, O.s.: Oryza sativa).

[0078]FIG. 42 illustrates the comparison of COQ-4 homologous proteins.

[0079]FIG. 43 illustrates Drosophila melanogaster COQ-4 protein (derived from gi|7293987|gb|AAF49344.1|CG3877).

[0080]FIG. 44 illustrates Homo sapiens COQ-4 protein (derived from gi|7705807|ref|NP_(—)057119.1|CGI-92).

[0081]FIG. 45 illustrates Schizosaccharomyces pombe COQ-4 protein (derived from gi|7493130|pir||T37755).

[0082]FIG. 46 illustrates Arabidopsis thaliana COQ-4 protein (derived from gi|4406761|gb|AAD20072.1|).

[0083]FIG. 47A illustrates part of Mus musculus COQ-4 protein (derived from gb|AA274683.1|AA274683); FIG. 47B part of Mus musculus COQ-4 protein (derived from dbj|AU051632.1|AU051632); FIG. 47C part of Mus musculus COQ-4 protein (derived from gb|AI157531.1|AI157531); and FIG. 47D Mus musculus COQ-4 consensus protein.

[0084]FIG. 48 illustrates Glycine max COQ-4 protein (derived from gb|AW201157.1|AW201157).

[0085]FIG. 49 illustrates Bos taurus COQ-4 protein (derived from gb|AW660771.1|AW660771).

[0086]FIG. 50 illustrates Medicago truncatula COQ-4 protein (derived from gb|AW696025.1|AW696025).

[0087]FIG. 51 illustrates Ancylostoma caninum COQ-4 protein (derived from gb|AW870537.1|AW870537).

[0088]FIG. 52 illustrates Trypanosoma cruzi COQ-4 Protein (derived from gb|AW330043.1|AW330043).

[0089]FIG. 53 illustrates Rattus rattus COQ-4 protein (derived from gb|AA800046.1|AA800046).

[0090]FIG. 54 illustrates Gossypium hirsutum COQ-4 protein (derived from gb|A|731097.1|AI731097).

[0091] FIGS. 55A-C illustrate the expression pattern of clk-2.

[0092] FIGS. 56A-E illustrate the telomere-lengthening phenotype of clk-2(qm37) mutants at different temperatures.

DETAILED DESCRIPTION OF THE INVENTION

[0093] For the first time, there is provided with the present invention a new method of increasing life span by modulating the biology of telomeres.

[0094] The Clk Phenotype of clk-2 Mutants

[0095] We had shown previously that clk-2 mutants have a phenotype similar to that of clk-1 mutants, including the maternal rescue effect, their slow development and behavior, and their increased life span (Hekimi, et al., Genetics 141, 1351 (1995); Lakowski, B. and Hekimi, S. Science 272, 1010 (1996). We have characterized the defects of clk-2 mutants in much further detail, the results of which follow. From 15° C. to 20° C. the phenotype of clk-2 mutants is similar to that of clk-1 mutants. The average developmental, reproductive and behavioral rates are dramatically slower, and the mean and maximum life span longer, than those of the wild type as summarized in Table 1. In particular, the embryonic development of clk-2(qm37) mutants lasts 17.0±1.5 hours (n=97) at 20° C., while the wild type lasts 13.2±0.7 hours (n=80). The post-embryonic development of clk-2 (qm37) mutants is also slower lasting 95.7±1.3 hours at 20° C. (n=73), while the wild-type worms take only 53.6±8.7 hours (n=184).

[0096] The defecation cycles are slowed down as well, occurring every 105.7±15.2 seconds in clk-2 mutants at 20° C. (n=10) and every 54.9±0.6 seconds in the wild type (n=70). The pumping rate is lower, 180.9±24.8 pumps per minute occurring in the clk-2 mutants at 20° C. (n=25), and 265.3±64.4 pumps per minute in the wild type (n=25). TABLE 1 Phenotypic characterization of clk-2(qm37) animals at 20° C. Maternally Wild Type rescue clk- (N2) clk-2(qm37) 2(qm37) Embryonic 13.2 ± 0.7 17.0 ± 1.5 13.3 ± 1.6 Development n = 80 n = 97 n = 40 (hours) Post-embryonic 53.6 ± 8.7 95.7 ± 1.3  53.9 ± 12.4 Development  n = 184 n = 73 n = 98 (hours) Self-brood 302.4 ± 30.5 83.4 113.9 ± 30.3 Size n = 20 n = 10 n = 24 (eggs) Peak Egg- 5.3 1.3  3.6 ± 0.9 laying Rate n = 10 n = 10 n = 24 (eggs per hour) Defecation 54.9 ± 0.6 105.7 ± 15.2 60.3 ± 9.0 (seconds) n = 70 n = 10 n = 8  Pumping 265.3 ± 64.4 180.9 ± 24.8 245.2 ± 24.6 (pumps per n = 25 n = 25 n = 11 minute)

[0097] In addition, we have also examined the self-brood size at 20° C. and found that is reduced in clk-2 mutants where it is 83.4 (n=10), while it is 302.4±30.5 in the wild type (n=20). The peak egg-laying rate is 1.3 (n=10) in clk-2 mutants at 20° C., and 5.3 (n=10) in the wild type. We have also examined the life span. clk-2(qm37) mutants live longer than the wild type, living on average 22.4±7.4 days (n=100) at 20° C. and having a maximum life span of 40 days, which is longer that the average life span of 19.3±5.3 days (n=100) and maximum life span of 32 days of wild-type N2 worms.

[0098] The developmental and behavioral phenotypes are fully maternally rescued, that is to say that homozygous clk-2/clk-2 mutants derived from a clk-2(qm37)/+heterozygous mother display wild-type phenotypes. In fact, the embryonic development of homozygous mutants derived from a heterozygous mother takes only 13.3±1.6 hours (n=40) and their post-embryonic development lasts only 53.9±12.4 hours (n=98) at 20° C. Also maternally rescued are both defecation, which occurs every 60.3±9.1 seconds at 20° C. (n=8) and pumping, which occurs at a rate of 245.2±24.6 pumps per minute at 20 C. (n=11). However, the reproductive phenotypes are only partially rescued by a wild-type copy of the gene clk-2 in the mother. The self-brood size is 113.9±30.3 at 20° C. (n=24), and the peak egg-laying rate is 3.6±0.9 (n=24). This indicates that the wild-type clk-2 gene in the mother induces an epigenetic state that lasts for only one generation. Erasure of the epigenetic state in the germ-line prevents the animal from having a wild-type rate of reproduction. In addition, the life span of maternally rescued homozygous mutants is dramatically shortened vs. both the mutant and the wild-type life span. Indeed, homozygous mutants derived from a heterozygous mother live only 14.9±4.1 days on average (n=106) and have a maximum life span of 27 days at 20° C. Interestingly, wild-type siblings of maternally rescued clk-2 live slightly shorter than wild-type N2 worms, 17.3±4.1 days (n=206). This observation indicates that wild-type physiological rates imposed by a maternal epigenetic setting are deleterious to animals that are partially incapable of regulating their physiological rates in response to environmental conditions. TABLE 2 Life span of mutants and double mutant combinations at 20° C. indicated in days Maximum Life Genotype Mean Life Span Span Wild type (N2) 19.3 ± 5.3 32 n = 100 clk-2(qm37) 22.4 ± 7.4 40 n = 100 Maternally 14.9 ± 4.1 27 rescued n = 106 clk-2 (qm37) Wild type (N2) 18.4 ± 4.6 31 n = 260 clk-2(qm37) 22.9 ± 7.3 45 n = 260 daf-16(m26) 18.1 ± 2.6 25 n = 260 daf-16(m26) clk- 21.7 ± 5.8 41 2(qm37) n = 260 daf-2 (e1370)  29.3 ± 10.3 51 n = 50 daf-2(e1370) clk-  54.5 ± 21.4 101  2(qm37) n = 50 eat-2 (ad465)  30.0 ± 7.0 42 n = 34 eat-2(ad465) clk- 26.6 ± 6.3 45 2(qm37) n = 50

[0099] We characterized the life span increase produced by clk-2 (qm37) by comparing it to that produced by other aging genes as summarized in table 2. Among the other genes that affect life span in worms, the best understood are the daf genes. Mutations in the eat genes prolong life span through caloric restriction by reducing the food intake of the animals, a process that also prolongs life span in vertebrates. Mutations in daf genes prolong life span by partial activation of the dauer formation pathway. The dauer stage is a dormant, long-lived, alternative developmental stage which is induced by adverse environmental conditions. The increased life span of all dauer formation mutants that have been tested is suppressed by loss of function mutations in daf-16.

[0100] In fact, we found that while daf-16(m26) lives 18.1±2.6 days on average with a maximum life span of 25 days, the double mutants daf-16(m26) clk-2(qm37) lives an average life span of 21.7±5.8 days with a maximum life span of 41 days. Furthermore, although double mutants with two long-lived dauer formation mutations do not live longer than mutants carrying only one of the component mutations, daf-2(e1370) clk-2(qm37) double mutants live substantially longer than daf-2, almost three times longer than the wild type. We have shown that while daf-2(e1370) lives 29.3±10.3 days on average with a maximum life span of 51 days, the double mutants daf-2(e1370) clk-2(qm37) lives an average life span of 54.5±21.4 days with a maximum life span of 101 days. In contrast to these observations, the effects of clk-2 and eat-2 are not additive. In fact, the double mutants live somewhat shorter than eat-2 mutants. We have shown that eat-2(ad465) lives 30.0±7.0 days on average with a maximum life span of 42 days, and that the double mutants daf-2(e1370) clk-2(qm37) live 26.6±6.3 days on average with a maximum life span of 45 days. These observations are also consistent with the finding that daf-2 eat-2 double mutants live longer than daf-2 or eat-2 mutants in isolation (Lakowski, B. and Hekimi, S. Science 272, 1010 (1996)). Together, these results show that daf-2 and clk-2 prolong life span by distinct mechanisms but that clk-2 works in a way that resembles caloric restriction.

[0101] The Strict Maternal Effect of the clk-2(qm37) Mutation

[0102] In addition to the Clk phenotype displayed by clk-2(qm37) mutants, they exhibit a temperature-sensitive embryonic lethal and sterile phenotypes at 25° C. We knew that qm37 is a temperature sensitive mutation and that the mutants lay dead embryos when they are transferred to 25° C. (Hekimi, S. et al., Genetics 141, 1351 (1995)). These findings have now been extended, and the phenotype of clk-2 mutants at 25° C. has been examined after a number of temperature shift experiments at different stages of development, from permissive to restrictive temperature and vice versa.

[0103] At the permissive temperatures (15 to 20° C.), clk-2 embryos all develop normally and grow up to become long-lived adults. However, when hermaphrodites that have developed at a permissive temperature are transferred to 25° C. before egg-laying begins, they produce only progeny that dies during embryogenesis at various stages of development. When these hermaphrodites, that have been producing dead embryos at 25° C., are transferred back to 18° C., they lay only dead eggs at first, but start to lay live eggs that develop into adults after having been 5-6 hours at 18° C. When hermaphrodites that are kept at 18° C., and that lay only live eggs, are transferred to 25° C. it also takes 5-6 hours before they lay only dead eggs. Both conditions (laying live or dead progeny) are fully reversible upon temperature shift even when the animal's entire post-embryonic development was carried out at a single temperature (permissive or non-permissive). In addition, when larvae that developed at the permissive temperature are shifted to 25° C., some arrest development and others reach a sterile and sick adulthood. These phenotypes are fully reversible as well. Finally, all these lethality and sterility phenotypes displayed by clk-2(qm37) mutants at 25° C. can be fully maternally rescued: heterozygous animals produce only live progeny at any temperature.

[0104] We have also found that the embryonic lethality at 25° C. is a strict maternal phenotype. That is to say that despite qm37 behaving as a recessive mutation, a wild-type allele in the genome of the embryo is not sufficient for survival if the mother was clk-2/clk-2 homozygous mutant. When clk-2 hermaphrodites are mated to wild-type males at 25° C. they nonetheless produce only dead embryos. When shifted to 18° C. at various times after mating they produce live males, indicating that the mating was successful. The strictly maternal lethal action of clk-2 indicates a very early focus of action, before activation of the zygotic genome.

[0105] To establish how early clk-2 acts during the development of the worm, we dissected embryos at the 2-4 cell stage from wild-type N2 and clk-2 mutant hermaphrodites kept at either permissive (20° C.) or non-permissive (25° C.) temperature and transferred them to the other temperature (or not, as a control). As summarized in table 3, we found that when development up to the 2-4 cell stage proceeded at the permissive temperature, almost all eggs hatched and carried out further embryonic and post-embryonic development at 20° C. {100% of dissected N2 eggs (n=35) hatched and 87% of dissected clk-2 eggs hatched (n=91)} or 25° C. {97% of dissected N2 eggs (n=36) hatched and 91% of dissected clk-2 eggs hatched (n=93)}. In contrast, when eggs had carried out development up to the 2-4 cell stage at 25° C. and were then transferred to 20° C., only very few clk-2 eggs hatched and succeeded in completing development at 20° C. {12% of dissected clk-2 eggs hatched (n=136)}. As a control, when N2 eggs had carried out development up to the 2-4 cell stage at 25° C. and were then transferred to 20° C., almost all hatched and succeeded in completing development at 20° C. {98%, n=45}, or at 25° C. {96%, n=45}. These results indicate that clk-2 is required for viability before the 2-4 cell stage. clk-2 is required in a narrow window between the very end of oogenesis and the initiation of embryonic development. TABLE 3 Survival of eggs at the 2-4 cell stage, dissected from mothers raised at 20 or 25° C. and transferred or not to another temperature % of eggs that % of eggs that hatch when hatch when developing at developing at Mothers 20° C. 25° C. N2 at 20° C. 100  97 n = 35 n = 36 clk-2 at 20° C. 87  91* n = 91 n = 93 N2 at 25° C. 98 96 n = 45 n = 45 clk-2 at 25° C.  12* n.d. n = 136

[0106] Indeed, clk-2 hermaphrodites that have spent 26 hours of adulthood at 25° C., carry on average 9.9 developing eggs in the uterus (n=125), but produce on average 10.7 dead eggs (n=133) when shifted down to permissive temperature. This observation indicates that, upon transfer from the lethal temperature, only one oocyte or embryo dies on average in addition to those that have already formed an eggshell. This corresponds to the time at which fertilization, oocyte meiosis, pronuclear formation and eggshell formation occurs. We observed early embryonic development using DIC microscopy but did not detect any obvious abnormality in the events which follow fertilization. The early embryos look invariably normal and healthy with cells and nuclei of normal size and shape. We also visualized DNA using Dapi in oocyte and early embryos and did not detect abnormal patterns of chromosome segregation or any other defects. Finally, meiosis per se is not affected as clk-2 homozygous males can sire abundant cross-progeny at 25° C. when mated to wild-type hermaphrodites.

[0107] clk-2 Positional Cloning, Gene Structure and Operon

[0108] We have molecularly identified the gene clk-2 by positional cloning. The gene was localized on the genetic map within an interval of 0.84 cM on the left cluster of linkage group III of Caenorhabditis elegans, between the genetic markers sma-4 and mab-5 (Hekimi, S. et al., Genetics 141, 1351 (1995)). We refined this genetic position by a series of additional mapping experiments involving the genetic markers sma-3, unc-36, lin-13, and lin-39 by multi- and two-point crosses. The following multi-point results were obtained (the genotypes whose progeny was scored is given in brackets): dpy-17 14 clk-2 18 unc-32 (clk-2/dpy-17 unc-32); lon-1 47 clk-2 23 unc-36 (clk-2/lon-1 unc-36); sma-4 35 clk-2 3 mab-5 14 unc-36 (clk-2/sma-4 mab-5 unc-36); sma-3 18 clk-2 0 lin-13 10 unc-36 (sma-3 clk-2 unc-36/lin-13); clk-2 3 lin-13 49 unc-32 (lin-13/clk-2 unc-32); sma-3 40 lin-39 0 clk-2 33 unc-36 (sma-3 clk-2 unc-36/lin-39). In addition, a two-point cross was carried out (clk-2 unc-36/++) and 5/630 Uncs were found to develop quickly (p=0.4 cM). We also found that the deletion nDf2O does not delete clk-2 and that the duplication qDp3 does include clk-2. We thus placed the gene clk-2 within an interval of 0.3 cM, between sma-3 (at −0.9 cM on LGIII) and lin-13 (at −0.6 cM on LGIII), and lying very close to the gene lin-39 (at −0.65 cM).

[0109] By aligning the genetic and physical maps, we predicted the physical region which likely would contain the clk-2 gene. Groups of cosmids from this region were tested for their ability to rescue the clk-2 mutant by DNA microinjection. clk-2 was rescued by a pool of 4 cosmids (H14A12, K07D8, C34A5, C07H6). Individual injection of cosmids C07H6 and C34A5 also rescued the clk-2 phenotype, narrowing the physical position of clk-2 to within approximately 15 kb. Fragments of cosmid C07H6 (obtained by restriction digests from base pair 31,528 to base pair 36,545 of cosmid C07H6 [Accession: AC006605]) were then tested for rescue and a short region of approximately 5 kb was shown to fully rescue the phenotype, indicating that this 5 kb fragment contains the clk-2 gene.

[0110] The identity of the gene was further confirmed by phenocopying the clk-2 phenotype with RNA interference (RNAi) experiments, that is the injection of double stranded RNA corresponding to the coding mRNA sequence of a gene of interest to fully abolish the function of this gene. Double stranded RNA was produced by in vitro transcription from a cDNA (EST 447b4, gift of Y. Kohara) that mapped to this region, and injected into wild-type as well as into clk-2(qm37) worms. All wild-type and clk-2 animals injected with clk-2 dsRNA initially produced embryos that hatched and developed into worms phenotypically resembling clk-2(qm37), that is, slow development, slow defecation and sterility. After 24 hours, the injected animals started laying only dead eggs. These results confirmed the identity of clk-2. The observation that RNAi-treated mothers produce dead eggs, a phenotype more severe than the weak embryonic lethality normally present in the clk-2(qm37) strain, indicated that qm37 is a partial loss-of-function mutation that displays the null phenotype only at 25° C. We further confirmed the identity of the gene by characterizing the molecular lesion underlying the clk-2 mutation. Genomic DNA from the clk-2(qm37) strain was isolated and the nucleotide sequence of the clk-2 region determined. The qm37 mutation is a G->A transition at in base 2321 of the cDNA.

[0111] The structure of the gene was established experimentally by determining the nucleotide sequence of the EST yk447b4 cDNA, thus defining the actual intron/exon boundaries in vivo and allowing to predict the encoded protein. The gene clk-2 is SL2 transpliced. We have further established the gene structure by RT-PCR experiments, which not only showed that clk-2 is SL2 transpliced, but also that the gene just upstream to clk-2, which we called cex-7, is expressed and is SL1 transpliced. The transplicing by SL1 of a gene placed upstream, and by SL2 of a gene downstream constitutes a hallmark of genes which are in an operon, and are transcriptionally co-expressed. Therefore, clk-2 and cex-7 are transcriptionally co-expressed, and thus play functionally related roles. The cDNA (yk215f6) that corresponds to cex-7 was also sequenced. The gene cex-7 encodes a predicted protein of 481 amino acid residues in length (FIG. 34), that is similar to a human polypeptide of 550 amino acids (FIG. 35).

[0112] clk-2 encodes a predicted protein of 877 amino acids and the clk-2(qm37) mutation is a cysteine to tyrosine substitution at residue 772 of the predicted protein. We have been able to detect the expressed protein by western blot analysis of protein extracted from both mutant and wild-type worms at different temperatures. CLK-2 is similar to unique predicted proteins in human (FIG. 3), Drosophila (FIG. 13), rice (FIG. 19), soybean (FIGS. 26-30) and to Saccharomyces cerevisiae Tel2p (FIG. 32) and in other species (FIGS. 7-12, 14, 17-19). The structural conservation among these proteins is illustrated by the alignment presented in FIGS. 38, 39, 40 and 41. No homologue of Tel2p had previously been recognized because aligning multiple sequences is necessary to reveal the homology. Tel2p has been shown to bind yeast telomeric DNA in a sequence-specific manner (Kota, R. S. Runge, K. W. Chromosoma 108, 278 (1999); Kota, R. S., Runge, K. W. Nucleic Acids Research 26, 1528 (1998)) and to affect the length of telomeres.

[0113] Expression Pattern of clk-2

[0114] We determined the spatial and temporal expression pattern of the gene clk-2 by analyzing transcript and protein levels (FIG. 55) and by examining transgenic worms carrying reporter fusions. Panel A of FIG. 55 illustrates Northern and Western (37) analyses of clk-2 at all developmental stages. The level of ck-2 mRNA appears uniform throughout pre-adult development (E, embryos; L1-L4, larval stages; A, adult; glp-4, adult glp-4 (bn2ts) mutants at 25° C.). The low level of clk-2 expression in L4 larvae and in glp-4 mutants that lack a germline at 25° C. suggest that most clk-2 RNA in adults is located in gametes. In contrast to the finding with mRNA, the level of CLK-2 protein is similar at all stages including adults (lower panel of A). Panel B of FIG. 55, clk-2 mRNA and protein levels (lower panel) in mutant backgrounds (glp-4 (bn2ts), fem-3 (q20ts), which produces only sperm at 25° C., and fem-2 (b245ts), which produces only oocytes at 25° C.). The mRNA and protein levels of clk-2 expression are similar to the wild type in fem-3 and elevated in fem-2 mutants. glp-4 mutants have wild type protein levels but reduced mRNA levels. clk-2 mRNA appears strongly elevated in clk-2 mutants. Panel C of FIG. 55, CLK-2 protein levels in wild type and clk-2 mutants at three temperatures. clk-2(qm37) is a missense (C772Y) and temperature-sensitive mutation. The level of CLK-2 is greatly reduced in the mutant, but does not change as a function of temperature in either the wild type or the mutant. Worms were raised at 20° C. except when specified otherwise.

[0115] We grew populations of worms synchronized at different developmental stages and extracted total or polyA+ selected RNA from them. The highest level of clk-2 mRNA is detected in young adults. We used several mutants to determine the origin of the transcript level in young adults. Since clk-2 mRNA level is highly reduced in glp-4(bn2ts) mutants that do not develop a germline at the non-permissive temperature, most of the RNA present in wild-type young adults is in the germline. Given the low abundance of RNA in L4 larvae which possess an already large germline but only a few male gametes, most of the clk-2 mRNA in wild-type adults is localized to meiotic gametes, in particular to oocytes.

[0116] We have analyzed the CLK-2 protein level in different genetic backgrounds and in worms grown at different temperatures. We immunodetected CLK-2 protein on western blots by using two different polyclonal antibodies, MG19 and MG20. We obtained these antibodies by injecting rabbits with a bacterially expressed His₁₀-CLK-2 protein. We found that the content of CLK-2 protein is uniform across developmental stages in wild type and in clk-2 animals. Furthermore, the concentration of CLK-2 is not different from the wild type in, glp-4 mutants which have no germline, nor in fem-3 and fem-2 mutants that contain only sperm and only oocytes, respectively. Taken together these results indicate that gametes specifically accumulate high levels of clk-2 mRNA, presumably as a store to be used by the embryo. Finally, we observed that in qm37 mutants, while the level of clk-2 mRNA appears slightly elevated, the level of CLK-2 protein is greatly reduced.

[0117] We constructed three reporter constructs of the clk-2 gene that comprised different upstream promoter regions and/or the coding region of the clk-2 gene fused to the green fluorescent protein. Two of the constructs are transcriptional fusions, one containing bases 36932 to 37319 and the other containing bases 36932 to 40010 of cosmid C07H6 [Accession: AC006605]. A third reporter construct (pMQ251) is a translational fusion that contains bases 30501 to 37319, except bases 35078 to 36545 which are part of the gene cex-7. We microinjected these reporter genes into wild type and clk-2(qm37) mutant worms, and analyzed numerous worms from several transgenic lines carrying these reporters. We observed that the clk-2 promoter region directs expression in all somatic tissues, including hypodermis, muscles, neurons, excretory system, gut, pharynx, somatic gonad, vulva, and presumably all cells. No expression was visible in the germline, despite the use of both standard and complex array mixes. This is commonly the case for transgenes in C. elegans and does not indicate an absence of expression in the germline tissue. A full length fusion protein between CLK-2 and GFP (encoded by the construct pMQ251) that complements the mutant phenotype for development, behavior and viability at 25° C., is localized exclusively into the cytoplasm, which is consistent with the absence of an obvious nuclear localization signal in the predicted protein. The pattern observed is not a consequence of overexpression as very small transgene concentrations have been used in complex arrays (Kelly et al., Genetics 146:227-238, 1997). However, although the nucleus appears dark in the fluorescent images, it still may contains very small amounts of the fusion protein. This analysis of expression indicates that CLK-2 protein is indeed produced in the nematode, as shown by western analysis on total C. elegans extracts using anti-CLK-2 antibodies.

[0118] Yeast Tel2p has been found to bind telomeric repeats in vitro, and thus is expected to be nuclear in vivo. However, it was found that CLK-2::GFP is excluded from the nucleus. Subtelomeric silencing and telomere length regulation can also be affected by events in the cytosol. For example, Hst2p, a cytosolic NAD+-dependent deacetylase homologous to Sir2p, can modulate nucleolar and telomeric silencing in yeast Perrod et al., EMBO J., 20(Nos 1 & 2), 197-209, 2001), and the nonsense-mediated mRNA decay pathway appears to affect both telomeric silencing and telomere length regulation (Lew et al., Molecular and Cellular Biology, 18(10):6121-6130, 1998). Other proteins that affect telomere length, like tankyrase Smith, S. and De Lange, Titia, J. of cell Science, 112:3649-3656, 1999), are mostly extranuclear Chi, N.-W., and Lodish, H. F., J. of Biological Chemistry, 275(49):38437-38444, 2000), with only a very small amount of protein localized to the telomeres Smith et al., Science 282:1484-1487, 1998).

[0119] The Role of clk-2

[0120] Telomere function has been found to affect replicative life span in yeast and in vertebrate cells. It also has also been shown to affect the immortality of the germline in C. elegans. However, an involvement of telomere function in determining the life span of muiticellular organisms has not been established prior to this work. Here we have shown that the maternal-effect clk-2 gene of C. elegans regulates telomere length, and prolongs life span by a mechanism that is distinct from the regulation of dauer formation but resembles caloric restriction, and encodes a protein that is similar to the yeast telomere binding protein Tel2p.

[0121] The timing of the lethal action of clk-2 (qm37) indicates a function for clk-2 during the events that immediately follow fertilization, including oocyte meiosis, pronuclei formation and karyogamy, and this would be consistent with the known importance of telomeres in meiosis. However, our examination of the morphology of chromosomes in oocytes and early embryos did not reveal any abnormalities. Similarly, although telomere function appears linked to double strand break repair and chromosome stability, including in worms, clk-2 mutants appear only moderately sensitive to ionizing radiation and do not display signs of chromosome instability. In fact, we examined the response of clk-2 (qm37) mutants to gamma-radiation and found that among the progeny of irradiated animals, the proportion of dead eggs and larvae was about 10 times higher than among the progeny of irradiated wild-type animals. There is also no report of a function of Tel2p in the response to ionizing radiation in yeast.

[0122] The null phenotype of tel2 is lethal but a hypomorphic mutation of tel2 results in short telomeres and slow growth (Runge, K. W. and Zakian, V. A. Molecular & Cellular Biology 16, 3094 (1996). Tel2p has been shown to be involved in telomere position effect (TPE) and thus contributes to silencing of sub-telomeric regions (Runge, K. W. and Zakian, V. A. Molecular & Cellular Biology 16, 3094 (1996), one of the best studied examples of epigenesis. Mutations in other genes, such as tell, that also result in telomere shortening do not result in abnormal TPE, indicating that the TPE defect in tel2 mutants is not a simple consequence of short telomeres. Furthermore, the rapid death and abnormal cellular morphology of cells fully lacking Tel2p suggests that Tel2p, like Rap1p and the Sir proteins, also functions at non-telomeric sites (Zakian, V. A. Ann. Rev. Genet. 30, 141 (1996)). In light of this, the absolute requirement for maternal clk-2 in embryogenesis suggests a function for CLK-2 in silencing genes that-are needed during some part of the worm's life cycle but that are deleterious when expressed during early development. The study of the mes genes which are required for the specification of the germline in C. elegans and can confer maternal-effect sterile phenotype has shown that mechanisms of silencing are part of the normal development of worms. Indeed, some of the mes genes have been found to encode proteins that resemble Polycomb group proteins and appear generally to be involved in the regulation of chromatin structure.

[0123] Mutations in clk-1 and clk-2(qm37) at the permissive temperature confer a similar CLK phenotype and in particular an increase of life span of similar magnitude (Lakowski, B. and Hekimi, S. Science 272, 1010 (1996) and show similar pattern of interactions with other aging genes (Lakowski, B. Hekimi, S. Proc. Nat. Acad. Sci. US 95, 13091 (1998)). CLK-1 is a mitochondrial protein of unknown function (Felkai, S. et al, EMBO Journal 18, 1783 (1999).). In an attempt to explain many puzzling features of the clk-1 phenotype, including the maternal effect, we have suggested that the action of CLK-1 is to indirectly, but specifically, regulate nuclear gene expression (Branicky R, C. Benard, S. Hekimi, Bioessays 22:48, 2000). One possibility might be that CLK-2 might be one of the molecules that implements changes in gene expression in response to alteration of CLK-1 activity. clk-1 clk-2 double mutants have a phenotype that is more severe than either of the single mutants (Lakowski, B. and Hekimi, S. Science 272, 1010 (1996). However, the phenotype of a double mutants containing the null allele clk-1 (qm3O) is not more severe than a double mutant containing the much weaker allele clk-1 (e2519), in contrast to the situation with clk-3, for which double mutants with clk-l(qm30) are much more severe than with clk-1 (e2519) (Lakowski, B. and Hekimi, S. Science 272, 1010 (1996). These observations indicate that at least part of the activity of clk-1 requires clk-2. Furthermore, clk-1 clk-2 double mutant embryos resemble clk-1 mutant in that the interphases of the embyronic cell cycles are slowed down, but mitoses appear unaltered. This indicates that clk-2 as well as clk-1 is involved in determining the rate of cellular multiplication, and thus affects mechanisms which are known to lead to cancer when deregulated.

[0124] Telomere function has also been implicated in the replicative life span of yeast, where Sir proteins mediate silencing at the telomeres and the HM loci. When displaced from the telomeres by mutation or by shortage of telomeric DNA, part of the Sir complex can move to the nucleolus where its action appears to prolong replicative life span. These and other studies indicate that telomeres are a reserve compartment for silencing factors and participate in regulating silencing in other parts of the genome. It has been suggested that the effect on cellular senescence of expressing telomerase in cultured human cells might be mediated by an effect on silencing rather than by preventing chromosome erosion. Therefore, clk-2 must be involved in determining cellular senescence, including in vertebrates, and affect in this manner aging and diseases linked to cellular senescence such as cancer.

[0125] As mentioned earlier, CLK-2 is similar to predicted proteins in vertebrates and plants as well as to Saccharomyces cerevisiae Tel2p. Tel2p has been shown to bind yeast telomeric DNA in a sequence-specific manner, and to affect the length of telomeres. We found that clk-2 also affected the length of telomeres in worms (FIG. 56). In worms, genomic DNA hybridization to telomeric probes after restriction digestion with HinfI reveals the end fragments of the chromosomes carrying the telomeres, which appear as smears, as well as fragments carrying tracts of telomeric repeats that are internal to the chromosome, which appear as discrete bands. The regions where the telomeric smears are the most intense are indicated by stippled lines. Two lanes are shown for each genotype and each temperature.

[0126] The length of telomeres in wild-type and clk-2 mutants was examined by Southern blotting at three temperatures, including the lethal temperature. For 18 and 20° C., worms were grown for numerous generations at each temperature before DNA extraction. Since clk-2(qm37) is lethal at 25° C., mixed stage worms from 20° C. were transferred to and grown at 25° C. for 3-4 days. Genomic DNA was prepared, HinfI digested and separated on a 0.6% agarose gel at 1.2Vcm⁻¹. Southern blots were hybridized with gamma ³²P DATP end-labelled TTAGGCTTAGGCTTAGGCTTAGGCTTAGGCTTAGGCTTAGGCTTAGG oligo-nucleotide. Use of a second type of probe, made by direct incorporation of alpha ³²P DATP during PCR amplification of telomeric repeats from the plasmid cTel55X with primers T7 and SHP1617 (GAATAATGAGAATTTTCAGGC), gave identical results. The extrachromosomal array in MQ691 clk-2(qm37); qmEx159 contains a clone with the entire coding sequence of clk-2 as well as the promoter of the operon but excluding cux-7 (bases 37319 to 31528 of cosmid C07H6, except bases 36544 to 35077) and rescues clk-2 mutant phenotypes. In clk-2 mutants, telomeres are two to three times longer than in the wild type on average (FIG. 56). However, the chromosomes are of wild-type length in strain MQ691, which carries an extrachromsomal array expressing wild-type CLK-2 in a clk-2(qm37) chromosomal background (FIG. 56) indicating that the alteration of telomere length clk-2 (qm37) mutants is indeed due to abnormal function of clk-2 in these mutants.

[0127] The length of terminal telomeric fragments in the animals of the strain MQ691, which carries an extrachromosomal array (qmEx159) containing functional wild-type CLK-2 that rescues development and behavior at 25° C. in a clk-2(qm37) chromosomal background, was further analyzed. A similar clone containing the qm37 mutation fails to rescue the Clk-2 phenotypes. In MQ691 animals, the length of terminal telomeric fragments appear very similar to the wild-type, and even shorter, indicating that the lengthened telomere phenotype of qm37 mutants is rescued by the expression of clk-2(+). The telomere length of non-transgenic animals of the strain MQ931, derived from MQ691, which have lost the extrachromosomal array and thus again lack clk-2(+) has been further examined. The terminal telomeric repeats in this strain are long again. Thus, the lengthened telomere phenotype of clk-2(qm37) can be rescued by clk-2(+) and reverses back to mutant length after the loss of the transgene.

[0128] In C. elegans, tracks of numerous TTAGGC telomeric repeats are present at the ends of the 6 chromosomes (Wicky C., et al., Proc. Natl. Acad. Sci. USA, 93:8983-8988, 1996). In addition, numerous interstitial blocks of perfect and degenerate telomeric repeats are located more internally to the chromosomes (C. elegans II. Edited by Riddel D et al. Published by Plainview, N.Y.: Cold Spring Harbor Laboratory Press (1997), pp 56-59, Chapter 3). Analysis of genomic DNA after restriction digestion with a frequent cutter that does not cleave within the telomeric repeats (HinfI), electrophoresis, and hybridization to telomeric probes, reveals the telomere-carrying end fragments of the chromosomes (Wicky C., et al., Proc. Natl. Acad. Sci. USA, 93:8983-8988, 1996). Telomeres, and thus the restriction fragments containing them, are heterogeneous in size and appear as smears. On the other hand, restriction fragments carrying tracts of internal telomeric repeats are of fixed size and appear as discrete bands in the 0.5-3 kb range (Ahmed S, Hodgkin J. Nature, 403(6766):159-64, 2000; and Wicky C., et al., Proc. Natl. Acad. Sci. USA, 93:8983-8988, 1996). The quality of visualization of the length of telomeres in C. elegans with a hybridization probe that detects telomeric repeats is marred by the numerous internal repeats that also hybridize to the probe. In particular, they can mask the detection of the telomeres of chromosomes that have small HinfI terminal telomeric fragments. To further describe the telomere phenotype of clk-2(qm37) mutants, the length of individual telomeres has been characterized. The subtelomeric regions just adjacent to the terminal telomeric repeats share no sequence homology among the chromosomes (Wicky C., et al., Proc. Natl. Acad. Sci. USA, 93:8983-8988, 1996). Taking advantage of this sequence diversity, probes specific to particular telomeres were designed. The size of a given HinfI terminal fragment is related to the fixed distance between the most exterior HinfI site of the chromosome and the beginning of the telomeric repeats, and by the variable number of terminal telomeric repeats. Upon genomic DNA digestion with HinfI and Southern blotting with a probe specific to a particular telomere, the terminal fragments, which are heterogeneous in size, again appear as a smear. Detailed results obtained for two individual telomeres are illustrated in FIG. 56.

[0129] The length of the terminal fragment of the left telomere of chromosome X is ˜1 kb longer in qm37 than in the wild type, ranging from 2.4 to 4.2 kb and from 1.7 to 2.8 kb, respectively. This telomere is of wild-type length in MQ691, which carries the rescuing transgene, and lengthens again to the clk-2(qm37) values in the non-rescued MQ931 strain. The length of another terminal fragment (left telomere of chromome IV) is also ˜1 kb longer in qm37 than in the wild type, ranging from 2.2 to 3.9 kb and from 1.8 to 2.8 kb respectively. This telomere becomes shorter than the wild type in MQ691, ranging from 1.3 to 2 kb only. This telomere acquires the mutant length again after loss of the transgene in MQ931. Thus, the overexpression of clk-2 can shorten the tracks of telomeric repeats, but not at each telomere.

[0130] Identity of the Gene cog-4 and the cog-4 (qm143) Mutants

[0131] The gene COQ7/CAT5 of the yeast S. cerevisiae is the homologous gene to clk-1 (Ewbank, J. J. et al, Science 275, 980 (1997); PCT/CA97/00768). While Coq7p does not structurally resemble an enzyme, it is required for ubiquinone biosynthesis in yeast. A second gene, COQ4 (Marbois, B. N. and Clark, C. J Biol Chem, 271, 2995 (1996) (Accession: NP_(—)010490), that is also required for ubiquinone biosynthesis in yeast, does not code for an enzyme, and like COQ7, has no homologue in bacteria. We have generated a deletion mutant worm to describe the role of the gene coq-4 and its functional relationships with the clk genes we have identified and described, including clk-1, clk-2, and gro-1.

[0132] The gene coq-4 in C. elegans largely corresponds to the predicted gene T03F1.2 of the cosmid T03F1 (Accession U88169). It is localized on LGI, between unc-73 and unc-11. coq-4 is less than 100 kb away from the characterized gene, unc-73, and less than 40 kb away from the other characterized gene, unc-11. coq-4 is 843 bp long and has four exons. We experimentally established the structure of the gene coq-4 by sequencing a cDNA clone, yk140a2. A second gene, T03F1.3, which is highly similar to phosphoglycerate kinase (PGK), is 264 bp upstream of coq-4 and, as we have shown, forms an operon with coq-4 and is thus transcriptionally co-expressed. We showed that Coq-4 is in the same operon as T03F1.3 by RT-PCR, that coq-4 is SL2 trans-spliced and that T03F1.3 is SL1 trans-spliced.

[0133] We have generated a coq-4-(qm143) deletion mutant by carrying out PCR-based mutant screen following a large scale EMS mutagenesis wild-type worm. coq-4 (qm143) has a 1469 bp deletion, which starts from 44 bp downstream of T03F1-3, and ends 406 bp downstream of coq-4. The predicted gene downstream is 1521 bp away from coq-4 and 1115 bp away from the deletion. Therefore, coq-4 (qm143) is a null mutant and it does not affect the coding sequence of any gene other than coq-4.

[0134] The Phenotype of cog-4 Mutants

[0135] cog-4(qm143) is a non-strict maternal-effect lethal mutation. Most of the progeny, from a homozygous coq-4 hermaphrodite, dies during embryogenesis. Very few eggs hatch, and those which do hatch fail to complete development and die as young larvae. We have also shown that maternal cog-4 product is sufficient for homozygous coq-4 to develop normally until adulthood. However, homozygous coq-4 adult worms from a heterozygous hermaphrodite (coq-4/+) are paralytic and are defective in egg-laying. Moreover, coq-4 homozygous mutants can be mated by N2 males and produce progeny, which grow normally. Taken together, these results indicate that either maternal or zygotic coq-4 product is sufficient for coq-4 mutant to go through embryonic and post-embryonic development. coq-4 deletion (qm143) is kept as a balanced strain, coq-4(qm143)/unc-73(e936). We demonstrated that the phenotypes of the coq-4 mutants, in particular the sterility, can be rescued by an extrachromosomal wild-type copy of coq-4 DNA fragment.

[0136] Expression Pattern of cog-4

[0137] The spatial expression pattern of coq-4 was determined by using translational reporter fusion to the green fluorescence protein, containing 2.2 kb of upstream promoter region. These constructs were injected into both N2 and heterozygous coq-4 (coq-4/unc-73), and animals of several transgenic lines were examined. We found that a functional coq-4::gfp is expressed in the hypodermis, muscles, the gut, the excretory canal and embryos. In addition, we detected that the reporter fusion localizes to the mitochondria, in particular, in muscle cells.

[0138] While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains and as may be applied to the essential features hereinbefore set forth, and as follows in the scope of the appended claims.

1 52 1 2814 DNA C. elegans clk-2 cDNA 1 ctcaagatga atttacgaag tcgcctggta aatgccacgg aacgtgctgt gctttttcaa 60 attttcaaag atgtgcagaa tgatccggaa aagtacgaca atgcagttga ggcgatctgt 120 gaatcaatcg actattttgg caaatttttg accgatagcg agtatcttac acaaatcaag 180 ccgattcttg atacacagtg cccaacaaag tcgataattt gcttctcgaa atgtttgaca 240 aaagtgagca cagatataaa tactaccaca tttcgagatg tgatcaccat gctcgactgg 300 ttgaagtatg tcgttgaaaa atcgctgaca agtgctattt gtagcagtct gaaagttaaa 360 gaaactgatg tcagtgcagt tcagttgtat cgagaattcg catcagcatg ttcaaatatt 420 ccggagaaag tttcgaattg ttgtgcaaag gcattgtctg gcgagcatgt caaatatatc 480 aacacggtta agtggatatt caaaatgaat ctggtgcaag gaattcaaaa ggctatgctt 540 cttgctcacg acgacattgt aactgctgcc ccgttcactt cattctacgg atccggtggt 600 ccttatatga agactgtcgc agaaattatt tcatctggaa gaaacataga tatcaccaac 660 aaggatgggc ttctagttca aatgattgaa tggattggtt cactaaacaa ttttgattct 720 caatggcgtc ggatgatgtt tctcatcttc caagagccca catatcaggg aattcaagtt 780 catgaatcac tactgacaac attgttccta atttcgaaaa gtgaccaaat cttgaaacga 840 tgtatcgaag ccactgatct gactggaaca ctgaagcgtg tagtgatggt taagctcccg 900 tttcagcgag ttctcaaacg aaagaccatc gagattctta tcaattttgt ttatcgaact 960 aaggaacaat ttgccatcca gctattagag acttctgtga aaatctggag tgatctcaat 1020 tacgcaaaaa gtgctccaga atcacaagaa cgacacatag tcagaatgat attatacttg 1080 gttcatcttt tcagaacatg ttcttcaatc gattgggagt cactcttcct gaactctatg 1140 gatggagttc attgtcgaat gagcatgttg cctatgtacg tccaaagtgg tatttttgtt 1200 aatcaagcac tgtgcaagca agcgacaaag catcgatcga aaacgcacgg atcagatgag 1260 caacctccag agactctaga agaaaacaaa ttcgtttcaa gtgaagtggg aaaaatatgg 1320 tttgaagaga tgacgtcaat tttggaacat ggatttaatt cttctacagt gaaagattct 1380 gagcgagttc gagaaaccgc caacgaaata accaaagacg attcgggtga agaatttgaa 1440 gaaaccaatg cacagcgtct tcaaaacaac aaagattcgg cagcaatcac atcgaaaaac 1500 aatctacgtt tagattctga tgatgacgaa gactttcctg actatcaagt taatgaatca 1560 gaaaagatct tcaagaattt agaaattgga gaagaaccga aaaataaagt gacacctcca 1620 gcatacattg cagatgcttt cgaaatgcta ttagagaaag aaaaatacga ggtttttgaa 1680 gcagctttct ttaatattac gaatttgatc aatcgccggc caattggatt tccacaaatc 1740 gctgagaagt tgttcatccg aatcctccat cttcaaaaca attttggaac gcctaaattc 1800 aaggaaactg ttgatgaaat tgcagttgca tgtatcactc agcgtccaga aattgtccca 1860 tctgtagtgc gtctgatcat tgcaccaggt caaggtttca gtatcaaaca acgtcttctt 1920 cattacattc acaatgctgc tgatggaatg ggtgcattgg ataagaaact tgaagagtgt 1980 gtaatggcgc aacaattgag aattggtggt ccaacgttaa gtattattct tcatcgaact 2040 ataaacactg attatgacga tgaggatgaa gatccccaca gacttttagt tcctgaatgg 2100 cgacgaatgg tggatgctcg cattgctgca aataccagaa gaattggaac gacgcgagag 2160 ccaccaagag ccggagttgt caatcgtctc gcacaagctg ccaaatatat gttttatcct 2220 ttgttggttt tgccacgtgg tgagaacgca agtcttttgg gcaaggactc cgatctactc 2280 gcctcactca tcatggttgc atcgatggtt tatgtgagat gtggcgtatg tcctcaaatt 2340 catcgaatgt caagtgagct tatatcatat gcaacgcctc atcgattctc tgaaaacgca 2400 aaactacgga ctgcctgcat cattgcccat ctgaatgtga cgactttgtt gcctggagat 2460 cttatggatg aactgtttga tgtaccggca cttattggat ggtttgattg ggccaattct 2520 gtactggtta atgcatcttc atcacaattg gaaaaggata tgactcgcca gtttggtcat 2580 agtgttacaa aacaccttca acgttatcat ccagctgtac tgcaacacca agacgtttaa 2640 atagttacta ttcacttgtt ttccttcttt tcaagtactg tatcatttct tactatcttg 2700 ccaacacttt gatctctacc tcgttcactt cttgctttgc cacccgttga tatcacctgt 2760 ctcattcatt tatcagcatg ttcataatat caaaaataaa atcttatcaa atgt 2814 2 876 PRT C. elegans clk-2 protein 2 Met Asn Leu Arg Ser Arg Leu Val Asn Ala Thr Glu Arg Ala Val Leu 1 5 10 15 Phe Gln Ile Phe Lys Asp Val Gln Asn Asp Pro Glu Lys Tyr Asp Asn 20 25 30 Ala Val Glu Ala Ile Cys Glu Ser Ile Asp Tyr Phe Gly Lys Phe Leu 35 40 45 Thr Asp Ser Glu Tyr Leu Thr Gln Ile Lys Pro Ile Leu Asp Thr Gln 50 55 60 Cys Pro Thr Lys Ser Ile Ile Cys Phe Ser Lys Cys Leu Thr Lys Val 65 70 75 80 Ser Thr Asp Ile Asn Thr Thr Thr Phe Arg Asp Val Ile Thr Met Leu 85 90 95 Asp Trp Leu Lys Tyr Val Val Glu Lys Ser Leu Thr Ser Ala Ile Cys 100 105 110 Ser Ser Leu Lys Val Lys Glu Thr Asp Val Ser Ala Val Gln Leu Tyr 115 120 125 Arg Glu Phe Ala Ser Ala Cys Ser Asn Ile Pro Glu Lys Val Ser Asn 130 135 140 Cys Cys Ala Lys Ala Leu Ser Gly Glu His Val Lys Tyr Ile Asn Thr 145 150 155 160 Val Lys Trp Ile Phe Lys Met Asn Leu Val Gln Gly Ile Gln Lys Ala 165 170 175 Met Leu Leu Ala His Asp Asp Ile Val Thr Ala Ala Pro Phe Thr Ser 180 185 190 Phe Tyr Gly Ser Gly Gly Pro Tyr Met Lys Thr Val Ala Glu Ile Ile 195 200 205 Ser Ser Gly Arg Ile Asp Ile Thr Asn Lys Asp Gly Leu Leu Val Gln 210 215 220 Met Ile Glu Trp Ile Gly Ser Leu Asn Asn Phe Asp Ser Gln Trp Arg 225 230 235 240 Arg Met Met Phe Leu Ile Phe Gln Glu Pro Thr Tyr Gln Gly Ile Gln 245 250 255 Val His Glu Ser Leu Leu Thr Thr Leu Phe Leu Ile Ser Lys Ser Asp 260 265 270 Gln Ile Leu Lys Arg Cys Ile Glu Ala Thr Asp Leu Thr Gly Thr Leu 275 280 285 Lys Arg Val Val Met Val Lys Leu Pro Phe Gln Arg Val Leu Lys Arg 290 295 300 Lys Thr Ile Glu Ile Leu Ile Asn Phe Val Tyr Arg Thr Lys Glu Gln 305 310 315 320 Phe Ala Ile Gln Leu Leu Glu Thr Ser Val Lys Ile Trp Ser Asp Leu 325 330 335 Asn Tyr Ala Lys Ser Ala Pro Glu Ser Gln Glu Arg His Ile Val Arg 340 345 350 Met Ile Leu Tyr Leu Val His Leu Phe Arg Thr Cys Ser Ser Ile Asp 355 360 365 Trp Glu Ser Leu Phe Leu Asn Ser Met Asp Gly Val His Cys Arg Met 370 375 380 Ser Met Leu Pro Met Tyr Val Gln Ser Gly Ile Phe Val Asn Gln Ala 385 390 395 400 Leu Cys Lys Gln Ala Thr Lys His Arg Ser Lys Thr His Gly Ser Asp 405 410 415 Glu Gln Pro Pro Glu Thr Leu Glu Glu Asn Lys Phe Val Ser Ser Glu 420 425 430 Val Gly Lys Ile Trp Phe Glu Glu Met Thr Ser Ile Leu Glu His Gly 435 440 445 Phe Asn Ser Ser Thr Val Lys Asp Ser Glu Arg Val Arg Glu Thr Ala 450 455 460 Asn Glu Ile Thr Lys Asp Asp Ser Gly Glu Glu Phe Glu Glu Thr Asn 465 470 475 480 Ala Gln Arg Leu Gln Asn Asn Lys Asp Ser Ala Ala Ile Thr Ser Lys 485 490 495 Asn Asn Leu Arg Leu Asp Ser Asp Asp Asp Glu Asp Phe Pro Asp Tyr 500 505 510 Gln Val Asn Glu Ser Glu Lys Ile Phe Lys Asn Leu Glu Ile Gly Glu 515 520 525 Glu Pro Lys Asn Lys Val Thr Pro Pro Ala Tyr Ile Ala Asp Ala Phe 530 535 540 Glu Met Leu Leu Glu Lys Glu Lys Tyr Glu Val Phe Glu Ala Ala Phe 545 550 555 560 Phe Asn Ile Thr Asn Leu Ile Asn Arg Arg Pro Ile Gly Phe Pro Gln 565 570 575 Ile Ala Glu Lys Leu Phe Ile Arg Ile Leu His Leu Gln Asn Asn Phe 580 585 590 Gly Thr Pro Lys Phe Lys Glu Thr Val Asp Glu Ile Ala Val Ala Cys 595 600 605 Ile Thr Gln Arg Pro Glu Ile Val Pro Ser Val Val Arg Leu Ile Ile 610 615 620 Ala Pro Gly Gln Gly Phe Ser Ile Lys Gln Arg Leu Leu His Tyr Ile 625 630 635 640 His Asn Ala Ala Asp Gly Met Gly Ala Leu Asp Lys Lys Leu Glu Glu 645 650 655 Cys Val Met Ala Gln Gln Leu Arg Ile Gly Gly Pro Thr Leu Ser Ile 660 665 670 Ile Leu His Arg Thr Ile Asn Thr Asp Tyr Asp Asp Glu Asp Glu Asp 675 680 685 Pro His Arg Leu Leu Val Pro Glu Trp Arg Arg Met Val Asp Ala Arg 690 695 700 Ile Ala Ala Asn Thr Arg Arg Ile Gly Thr Thr Arg Glu Pro Pro Arg 705 710 715 720 Ala Gly Val Val Asn Arg Leu Ala Gln Ala Ala Lys Tyr Met Phe Tyr 725 730 735 Pro Leu Leu Val Leu Pro Arg Gly Glu Asn Ala Ser Leu Leu Gly Lys 740 745 750 Asp Ser Asp Leu Leu Ala Ser Leu Ile Met Val Ala Ser Met Val Tyr 755 760 765 Val Arg Cys Gly Val Cys Pro Gln Ile His Arg Met Ser Ser Glu Leu 770 775 780 Ile Ser Tyr Ala Thr Pro His Arg Phe Ser Glu Asn Ala Lys Leu Arg 785 790 795 800 Thr Ala Cys Ile Ile Ala His Leu Asn Val Thr Thr Leu Leu Pro Gly 805 810 815 Asp Leu Met Asp Glu Leu Phe Asp Val Pro Ala Leu Ile Gly Trp Phe 820 825 830 Asp Trp Ala Asn Ser Val Leu Val Asn Ala Ser Ser Ser Gln Leu Glu 835 840 845 Lys Asp Met Thr Arg Gln Phe Gly His Ser Val Thr Lys His Leu Gln 850 855 860 Arg His His Pro Ala Val Leu Gln His Gln Asp Val 865 870 875 3 836 PRT Homo sapiens clk-2 protein 3 Met Glu Pro Ala Pro Ser Glu Val Arg Leu Ala Val Arg Glu Ala Ile 1 5 10 15 His Ala Leu Ser Ser Ser Glu Asp Gly Gly His Ile Phe Cys Thr Leu 20 25 30 Glu Ser Leu Lys Arg Tyr Leu Gly Glu Met Glu Pro Pro Ala Leu Pro 35 40 45 Arg Glu Lys Glu Glu Phe Ala Ser Ala His Phe Ser Pro Val Leu Arg 50 55 60 Cys Leu Ala Ser Arg Leu Ser Pro Ala Trp Leu Glu Leu Leu Pro His 65 70 75 80 Gly Arg Leu Glu Glu Leu Trp Ala Ser Phe Phe Leu Glu Gly Pro Ala 85 90 95 Asp Gln Ala Phe Leu Val Leu Met Glu Thr Ile Glu Gly Ala Ala Gly 100 105 110 Pro Ser Phe Arg Leu Met Lys Met Ala Arg Leu Leu Ala Arg Phe Leu 115 120 125 Arg Glu Gly Arg Leu Ala Val Leu Met Glu Ala Gln Cys Arg Gln Gln 130 135 140 Thr Gln Pro Gly Phe Ile Leu Leu Arg Glu Thr Leu Leu Gly Lys Val 145 150 155 160 Val Ala Leu Pro Asp His Leu Gly Asn Arg Leu Gln Gln Glu Asn Leu 165 170 175 Ala Glu Phe Phe Pro Gln Asn Tyr Phe Arg Leu Leu Gly Glu Glu Val 180 185 190 Val Arg Val Leu Gln Ala Val Val Asp Ser Leu Gln Gly Gly Leu Asp 195 200 205 Ser Ser Val Ser Phe Val Ser Gln Val Leu Gly Lys Ala Cys Val His 210 215 220 Gly Arg Gln Gln Glu Ile Leu Gly Val Leu Val Pro Arg Leu Ala Ala 225 230 235 240 Leu Thr Gln Gly Ser Tyr Leu His Gln Arg Val Cys Trp Arg Leu Val 245 250 255 Glu Gln Val Pro Asp Arg Ala Met Glu Ala Val Leu Thr Gly Leu Val 260 265 270 Glu Ala Ala Leu Gly Pro Glu Val Leu Ser Arg Leu Leu Gly Asn Leu 275 280 285 Val Val Lys Asn Lys Lys Ala Gln Phe Val Met Thr Gln Lys Leu Leu 290 295 300 Phe Leu Gln Ser Arg Leu Thr Thr Pro Met Leu Gln Ser Leu Leu Gly 305 310 315 320 His Leu Ala Met Asp Ser Gln Arg Arg Pro Leu Leu Leu Gln Val Leu 325 330 335 Lys Glu Leu Leu Glu Thr Trp Gly Ser Ser Ser Ala Ile Arg His Thr 340 345 350 Pro Leu Pro Gln Gln Arg His Val Ser Lys Ala Val Leu Ile Cys Leu 355 360 365 Ala Gln Leu Gly Glu Pro Glu Leu Arg Asp Ser Arg Asp Glu Leu Leu 370 375 380 Ala Ser Met Met Ala Gly Val Lys Cys Arg Leu Asp Ser Ser Leu Pro 385 390 395 400 Pro Val Arg Arg Leu Gly Met Ile Val Ala Glu Val Val Ser Ala Arg 405 410 415 Ile His Pro Glu Gly Pro Pro Leu Lys Phe Gln Tyr Glu Glu Asp Glu 420 425 430 Leu Ser Leu Glu Leu Leu Ala Leu Ala Ser Pro Gln Pro Ala Gly Asp 435 440 445 Gly Ala Ser Glu Ala Gly Thr Ser Leu Val Pro Ala Thr Ala Glu Pro 450 455 460 Pro Ala Glu Thr Pro Ala Glu Ile Val Asp Gly Gly Val Pro Gln Ala 465 470 475 480 Gln Leu Ala Gly Ser Asp Ser Asp Leu Asp Ser Asp Asp Glu Phe Val 485 490 495 Pro Tyr Asp Met Ser Gly Asp Arg Glu Leu Lys Ser Ser Lys Ala Pro 500 505 510 Ala Tyr Val Arg Asp Cys Val Glu Ala Leu Thr Thr Ser Glu Asp Ile 515 520 525 Glu Arg Trp Glu Ala Ala Leu Arg Ala Leu Glu Gly Leu Val Tyr Arg 530 535 540 Ser Pro Thr Ala Thr Arg Glu Val Ser Val Glu Leu Ala Lys Val Leu 545 550 555 560 Leu His Leu Glu Glu Lys Thr Cys Val Val Gly Phe Ala Gly Leu Arg 565 570 575 Gln Arg Ala Leu Val Ala Val Thr Val Thr Asp Pro Ala Pro Val Ala 580 585 590 Asp Tyr Leu Thr Ser Gln Phe Tyr Ala Leu Asn Tyr Ser Leu Arg Gln 595 600 605 Arg Met Asp Ile Leu Asp Val Leu Thr Leu Ala Ala Gln Glu Leu Ser 610 615 620 Arg Pro Gly Cys Leu Gly Arg Thr Pro Gln Pro Gly Ser Pro Ser Pro 625 630 635 640 Asn Thr Pro Cys Leu Pro Glu Ala Ala Val Ser Gln Pro Gly Ser Ala 645 650 655 Val Ala Ser Asp Trp Arg Val Val Val Glu Glu Arg Ile Arg Ser Lys 660 665 670 Thr Gln Arg Leu Ser Lys Gly Gly Pro Arg Gln Gly Pro Ala Gly Ser 675 680 685 Pro Ser Arg Phe Asn Ser Val Ala Gly His Phe Phe Phe Pro Leu Leu 690 695 700 Gln Arg Phe Asp Arg Pro Leu Val Thr Phe Asp Leu Leu Gly Glu Asp 705 710 715 720 Gln Leu Val Leu Gly Arg Leu Ala His Thr Leu Gly Ala Leu Met Cys 725 730 735 Leu Ala Val Asn Thr Thr Val Ala Val Ala Met Gly Lys Ala Leu Leu 740 745 750 Glu Phe Val Trp Ala Leu Arg Phe His Ile Asp Ala Tyr Val Arg Gln 755 760 765 Gly Leu Leu Ser Ala Val Ser Ser Val Leu Leu Ser Leu Pro Ala Ala 770 775 780 Leu Leu Glu Asp Leu Met Asp Glu Leu Leu Glu Ala Arg Ser Trp Leu 785 790 795 800 Ala Asp Val Ala Glu Lys Asp Pro Asp Glu Asp Cys Arg Thr Leu Ala 805 810 815 Leu Arg Ala Leu Leu Leu Leu Gln Arg Leu Lys Asn Arg Leu Leu Pro 820 825 830 Pro Ala Ser Pro 835 4 3320 DNA Homo sapiens clk-2 4 gcgccccaga ggctcaagaa aacccgcggg agcctcgccc ggacccagga actcgtgctc 60 ggggccaacc ggctgggccg cgatcgcgtt tcgtccgggg ccgcggcggc cgtggggaat 120 cggctgcagc gaatcggtgg cgcgcggcgc ctgagcgcgc tgcagtcacc cgggagccgg 180 gtccaggtcg tcttcccgtg acgcccagat ctgtcctgca ggatggagcc agcaccctca 240 gaggttcgac tcgccgtccg ggaagccatt catgccctct cgtcttcgga ggatggcggc 300 cacatcttct gcaccctgga gtccctgaag cggtatctcg gtgagatgga gcctccagcg 360 ctcccgaggg agaaggagga gtttgcctcg gcccacttct cgcctgtcct cagatgtctt 420 gccagcaggc tgagcccagc ctggctggag ctgctgcccc atggccgcct ggaggagctg 480 tgggccagct tcttcctgga gggcccggcg gaccaagcct tcctggtgtt gatggagacc 540 atcgagggtg ctgcgggccc cagcttccgg ctgatgaaga tggcgcggct gctggccaga 600 ttcctgcgcg agggccggct ggcagtgctg atggaggcgc agtgtcggca gcagacgcgg 660 cccggcttca tcctgctccg ggagacgctg ctgggcaagg tggtggccct gcccgatcac 720 ctgggcaacc gcctgcagca ggagaacttg gccgagttct tcccccagaa ctacttccgc 780 ctgctcggcg aggaggtcgt ccgggtgctg caggcggttg tggactctct ccaaggtggc 840 ctggattcct ccgtgtcctt cgtgtctcag gtccttggga aagcctgtgt ccacgggagg 900 cagcaggaga tcctgggcgt gctggtaccc cggctggcag cgctcaccca gggcagctac 960 ctgcaccagc gcgtctgctg gcgcctggtg gagcaagtgc cggaccgggc catggaggct 1020 gtgctgaccg ggctggtgga ggccgcactg gggcctgagg tcctttcgag actgctgggg 1080 aacctggtgg tgaagaacaa gaaggcccag tttgtgatga cccagaagct tctgttctta 1140 cagtcccggc tcacgacgcc catgctgcag agcctgctgg gccatctggc catggacagc 1200 cagcggcgcc cgctcctgct gcaggtgctg aaggagctgt tggagacgtg gggcagcagc 1260 agtgccatcc gccacactcc cctgccgcag cagcgccacg tcagcaaggc tgtcctcatc 1320 tgcctggcgc aactcgggga gccggaactg cgggacagcc gggatgaact gctggccagc 1380 atgatggcgg gcgtgaagtg ccgcctggac agtagcctgc cccccgtgcg acgcctgggc 1440 atgatcgtgg cagaggtcgt tagtgcccgg atccaccccg aggggcctcc cctgaaattc 1500 cagtacgaag aggatgaact gagcctcgag ctgctggcct tggcctcccc ccagcctgcg 1560 ggtgacggcg cctcggaggc gggcacgtcc ctcgttccag ccacggcaga gccccctgca 1620 gagacccccg cagagatcgt ggatggcggc gtcccccaag cacagctggc gggctctgac 1680 tcggacctgg acagcgatga tgagtttgtc ccctacgaca tgtcggggga cagagagctg 1740 aagagcagca aggctcctgc ctacgtccgg gactgcgtgg aagccctgac cacgtctgag 1800 gacatagagc gctgggaggc agccctgcgg gcccttgagg gcctggtcta caggagcccc 1860 acagccactc gggaggtgag cgtggagctg gccaaggtgc ttctgcatct ggaggagaag 1920 acctgtgtgg tgggatttgc agggctgcgc cagagagccc tggtggccgt cacggtcaca 1980 gacccggccc cggtggccga ctatctgacc tcacagttct atgccctcaa ctacagcctc 2040 cggcagcgca tggacatcct ggatgtgctg actctggctg cccaggagct gtctaggcct 2100 gggtgcctcg ggaggactcc ccaacctggc tccccaagtc ccaacacccc gtgcctgcca 2160 gaggcagccg tctctcagcc tggcagtgcc gtggcgtctg actggcgggt ggtggtggag 2220 gagcggatca gaagcaagac ccagcggctc tccaagggtg gcccgaggca gggcccggca 2280 ggcagcccca gcagattcaa ctccgtggcc ggccacttct tcttccccct ccttcagcgc 2340 tttgacaggc ctctggtgac cttcgacctc ttgggagaag accagctggt tctcggaagg 2400 ctggcgcaca ccttaggggc cctgatgtgc ctggctgtta acaccacggt ggctgtggcc 2460 atgggcaagg ccctgctgga attcgtgtgg gcccttcgct tccacatcga tgcctacgtg 2520 cgccaggggc tgttgtcggc cgtctcctcc gtcctgctca gcctgcctgc tgcgcgcctg 2580 ctggaggacc tgatggacga gctgctggaa gcccggtcct ggctggcgga cgtggctgag 2640 aaagacccgg acgaggactg caggacgctg gcactgaggg ccctgctgct tctgcagaga 2700 ctcaagaaca ggctcctccc acccgcgtct ccctagtccc tggaggcctc cccaggacca 2760 ccctcgccga cagcaaggca ggcggctgag cagcggcctg gagcagcaga gccaggcttt 2820 gtagcgaggc caggtcttcg gccgcatccg gtacggagag tgcagatgca ggaaggcccg 2880 gcctgccgct atttatagtg cagccagtcc gctaaaaata cactgggcct gggcactgcc 2940 cgccgggaca tggcagcctg gacgtggggc tggggctgtg ggcgctgctg gcggggttga 3000 ctcttccagt gagggcagaa ccaggctggc aggaggggag gacggtgtac ctgctgctca 3060 gagcccccaa ggctctcctc tgagagccac caagcaggac agagcagctc ttgtcccagg 3120 tccctcgggc tgagcgccgt gtcaccagga gaatagtgct cacagcccag gcagggtgtg 3180 tggctcctgg atgggctcgt ggggcgggat gggacagggc acgggctctc agaaaataaa 3240 ctgctttatt ggaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 3300 aaaaaaaaaa aaaaaaaaaa 3320 5 466 DNA Mus musculus clk-2 cDNA 5 gtgacccgca agctcctgct cctgcagtac cagcacacga cacccatggt gcagagcctg 60 ttggggtact tggctctaga cagtcagcgg cggccactcc tcatacaggt gcttaaggag 120 ctgctggaga cctggggctg cagcagtgct gtccgacaca cacccctgga gcagcagtgt 180 tacatcagca aggccatcct tgtctgcctg gcacacctcg gggagccgga gctgcaggac 240 atccgggatg aattgctggc cagcatgatg gcaggcgtga agtgccgcct ggatagcagc 300 ctgccccctg tgcgccgctt gggcatgatt gtggccgagg tcatcagctc caggatccac 360 cctgaggggc ctctcctgaa attccaatat gaagatgacg agatgagccg tgagttgctg 420 gccttggcta ccccagagcc tgcgggtgac tgctcctcgg tgtcac 466 6 267 DNA Artificial Sequence part of Mus musculus cDNA 6 tgtcccatgt gctgacttct gaggtggctg tgctagtggg taaggccctg ctggagtttg 60 tatgggccct tcgcttccac gttgacattt acgtgcgccg gggcttgctg tctgctgtgt 120 cctctgtcct cctcagtgta cccacagagc ggctgctggg ggacctgcca gatgagctac 180 tggaagccag atcctggttg gcagatgtgg ctgagaagga tgtggacgag gactgtaggg 240 agctggcagt aagggctctg ctgcttc 267 7 337 DNA Artificial Sequence part of Mus musculus cDNA 7 tcacagttct atggcctaaa ctatagcctc cgccagcgca tggacatcct ggacgtcctt 60 gttctggctg cccaggcact gtctcggcca aagagcctgc agagacgttc ccagcacggt 120 ccccccgttc ctggcaccat gtgttcacca gcactagccg tttctcagac tggcaatgtc 180 gctgctcctg actggcaggt ggttgtggag gagcggatca gaagcaagac ccggaggttc 240 tcgaagggct gtcctcagcg ggacgtgtca ggcgtcccca atgaattcag ctctgtggct 300 ggctacttct tcttccccct ccttcagcac tttgaca 337 8 153 PRT unknown part of Mus musculus clk-2 protein 8 Val Thr Arg Lys Leu Leu Leu Leu Gln Tyr Gln His Thr Thr Pro Met 1 5 10 15 Val Gln Ser Leu Leu Gly Tyr Leu Ala Leu Asp Ser Gln Arg Arg Pro 20 25 30 Leu Leu Ile Gln Val Leu Lys Glu Leu Leu Glu Thr Trp Gly Cys Ser 35 40 45 Ser Ala Val Arg His Thr Pro Leu Glu Gln Gln Cys Tyr Ile Ser Lys 50 55 60 Ala Ile Leu Val Cys Leu Ala His Leu Gly Glu Pro Glu Leu Gln Asp 65 70 75 80 Ile Arg Asp Glu Leu Leu Ala Ser Met Met Ala Gly Val Lys Cys Arg 85 90 95 Leu Asp Ser Ser Leu Pro Pro Val Arg Arg Leu Gly Met Ile Val Ala 100 105 110 Glu Val Ile Ser Ser Arg Ile His Pro Glu Gly Pro Leu Leu Lys Phe 115 120 125 Gln Tyr Glu Asp Asp Glu Met Ser Arg Glu Leu Leu Ala Leu Ala Thr 130 135 140 Pro Glu Pro Ala Gly Asp Cys Ser Ser 145 150 9 112 PRT unknown part of Mus musculus clk-2 protein 9 Ser Gln Phe Tyr Gly Leu Asn Tyr Ser Leu Arg Gln Arg Met Asp Ile 1 5 10 15 Leu Asp Val Leu Val Leu Ala Ala Gln Ala Leu Ser Arg Pro Lys Ser 20 25 30 Leu Gln Arg Arg Ser Gln His Gly Pro Pro Val Pro Gly Thr Met Cys 35 40 45 Ser Pro Ala Leu Ala Val Ser Gln Thr Gly Asn Val Ala Ala Pro Asp 50 55 60 Trp Gln Val Val Val Glu Glu Arg Ile Arg Ser Lys Thr Arg Arg Phe 65 70 75 80 Ser Lys Gly Cys Pro Gln Arg Asp Val Ser Gly Val Pro Asn Glu Phe 85 90 95 Ser Ser Val Ala Gly Tyr Phe Phe Phe Pro Leu Leu Gln His Phe Asp 100 105 110 10 85 PRT unknown part of Mus musculus clk-2 protein 10 Leu Thr Ser Glu Val Ala Val Leu Val Gly Lys Ala Leu Leu Glu Phe 1 5 10 15 Val Trp Ala Leu Arg Phe His Val Asp Ile Tyr Val Arg Arg Gly Leu 20 25 30 Leu Ser Ala Val Ser Ser Val Leu Leu Ser Val Pro Thr Glu Arg Leu 35 40 45 Leu Gly Asp Leu Pro Asp Glu Leu Leu Glu Ala Arg Ser Trp Leu Ala 50 55 60 Asp Val Ala Glu Lys Asp Val Asp Glu Asp Cys Arg Glu Leu Ala Val 65 70 75 80 Arg Ala Leu Leu Leu 85 11 350 PRT unknown composite protein sequence of Mus musculus clk-2 11 Val Thr Arg Lys Leu Leu Leu Leu Gln Tyr Gln His Thr Thr Pro Met 1 5 10 15 Val Gln Ser Leu Leu Gly Tyr Leu Ala Leu Asp Ser Gln Arg Arg Pro 20 25 30 Leu Leu Ile Gln Val Leu Lys Glu Leu Leu Glu Thr Trp Gly Cys Ser 35 40 45 Ser Ala Val Arg His Thr Pro Leu Glu Gln Gln Cys Tyr Ile Ser Lys 50 55 60 Ala Ile Leu Val Cys Leu Ala His Leu Gly Glu Pro Glu Leu Gln Asp 65 70 75 80 Ile Arg Asp Glu Leu Leu Ala Ser Met Met Ala Gly Val Lys Cys Arg 85 90 95 Leu Asp Ser Ser Leu Pro Pro Val Arg Arg Leu Gly Met Ile Val Ala 100 105 110 Glu Val Ile Ser Ser Arg Ile His Pro Glu Gly Pro Leu Leu Lys Phe 115 120 125 Gln Tyr Glu Asp Asp Glu Met Ser Arg Glu Leu Leu Ala Leu Ala Thr 130 135 140 Pro Glu Pro Ala Gly Asp Cys Ser Ser Ser Gln Phe Tyr Gly Leu Asn 145 150 155 160 Tyr Ser Leu Arg Gln Arg Met Asp Ile Leu Asp Val Leu Val Leu Ala 165 170 175 Ala Gln Ala Leu Ser Arg Pro Lys Ser Leu Gln Arg Arg Ser Gln His 180 185 190 Gly Pro Pro Val Pro Gly Thr Met Cys Ser Pro Ala Leu Ala Val Ser 195 200 205 Gln Thr Gly Asn Val Ala Ala Pro Asp Trp Gln Val Val Val Glu Glu 210 215 220 Arg Ile Arg Ser Lys Thr Arg Arg Phe Ser Lys Gly Cys Pro Gln Arg 225 230 235 240 Asp Val Ser Gly Val Pro Asn Glu Phe Ser Ser Val Ala Gly Tyr Phe 245 250 255 Phe Phe Pro Leu Leu Gln His Phe Asp Leu Thr Ser Glu Val Ala Val 260 265 270 Leu Val Gly Lys Ala Leu Leu Glu Phe Val Trp Ala Leu Arg Phe His 275 280 285 Val Asp Ile Tyr Val Arg Arg Gly Leu Leu Ser Ala Val Ser Ser Val 290 295 300 Leu Leu Ser Val Pro Thr Glu Arg Leu Leu Gly Asp Leu Pro Asp Glu 305 310 315 320 Leu Leu Glu Ala Arg Ser Trp Leu Ala Asp Val Ala Glu Lys Asp Val 325 330 335 Asp Glu Asp Cys Arg Glu Leu Ala Val Arg Ala Leu Leu Leu 340 345 350 12 122 PRT unknown part of Sus Scrofa clk-2 protein 12 Lys Ala Pro Val Tyr Val Arg Asp Cys Val Glu Ala Leu Thr Ala Ser 1 5 10 15 Glu Asp Trp Glu Arg Trp Glu Ala Ala Leu Arg Ala Leu Glu Gly Leu 20 25 30 Val Phe Arg Ser Pro Ala Ala Ala Arg Glu Val Ser Val Glu Leu Ala 35 40 45 Lys Val Leu Leu His Leu Glu Glu Lys Thr Ala Val Ala Gly Phe Glu 50 55 60 Gly Leu Arg Gln Arg Ala Leu Val Ala Val Thr Val Thr Asp Pro Ala 65 70 75 80 Arg Val Ala Glu Tyr Leu Thr Ala Gln Phe Tyr Ala Leu Asn Tyr Ser 85 90 95 Leu Arg Gln Arg Met Asp Ile Leu Asp Val Leu Thr Leu Ala Ala Gln 100 105 110 Glu Leu Ser Arg Pro Gly Arg Leu Gly Arg 115 120 13 554 PRT D. melanogaster clk-2 13 Leu Ile Ser Leu Pro Ala Gln Val Ala Asn Arg Leu Gly Arg Arg Leu 1 5 10 15 Pro Glu Thr Phe Ala Pro Val Ser Tyr Gln Lys Leu Leu Leu Arg Gln 20 25 30 Trp Leu Lys Ser Leu His Phe Val Leu Gln Cys Asp Asp Asn Arg Glu 35 40 45 Tyr Phe Asp Leu Glu Pro Tyr Ser Trp Leu Leu Ser Gln Ala Ile Asn 50 55 60 Leu Ile Tyr Asp Val Ser Thr Leu Glu Ser Leu Leu Arg Val Leu Lys 65 70 75 80 Asp Tyr Ala Val Ala Pro Arg Gly Arg Lys Val Val His Thr Ile Leu 85 90 95 Lys Glu Leu Asp Pro Ala Ala Cys Leu Lys Thr Ala Gln Ser Ala Leu 100 105 110 Ser Ala Gly Leu Asn Leu Tyr Val Leu Ile Gly Ala Ala Thr Leu Glu 115 120 125 Thr Pro His Trp Lys His Cys Leu Leu Gln Lys Leu Pro Leu Gln Arg 130 135 140 Thr Pro Val Asp Asn Lys Gln Leu Ile Thr Leu Ala Ser Tyr Leu Asn 145 150 155 160 Ala Val Ala Pro Ala Gln Leu Gln Val Leu Leu Asn Gln Leu Leu Gly 165 170 175 Ile Trp Ser Lys Arg Ile Ser Leu Gln Lys Leu Gly Ser Gln Glu His 180 185 190 Leu Ala Ile Ser Lys Leu Leu Val Leu Ala Gln Leu Asp Ser Asp Asp 195 200 205 Asp Glu Pro Leu Asp Glu Asp Asp Asp Glu Leu Lys Pro Tyr Asp Met 210 215 220 Ser Asn Asp Thr Thr Thr Thr Ile Asp Gln Arg Pro Lys Phe Val Ile 225 230 235 240 Asp Leu Leu His Leu Leu Arg Glu Lys Val Glu Asn Tyr Gln Val Phe 245 250 255 Glu Gly Ala Leu Gly Thr Ala Glu Gln Leu Ile Arg Gly Gln Leu Ala 260 265 270 Lys His Asp Thr Gln Leu Ala Leu Asp Leu Leu Gln Leu Phe Leu Val 275 280 285 Met Glu Met Gln Phe Tyr Tyr Glu Gln Phe Glu Arg Thr Gln Phe Lys 290 295 300 Cys Cys Val Ala Ile Cys Val Ala His Pro Gly Pro Cys Ala Glu Tyr 305 310 315 320 Leu Cys Arg Gln Phe His Thr Asp Asn Ser Phe Tyr Ser Ala Ser Val 325 330 335 Arg Ile Leu Ile Leu Gln Val Leu Ala Ala Thr Ala Lys Glu Leu Ser 340 345 350 Gly Asp Glu Asn Met Gln Asn Glu Met Glu Ile Val Asp Val Ile Pro 355 360 365 Pro Ala Ala Lys His Pro Arg Lys Phe Glu Phe Gln Gln Glu Glu Glu 370 375 380 Ser Pro Ala Ala Arg Leu Ala Ala Ala Gln Arg Ile Ile Arg Asp Arg 385 390 395 400 Leu Arg Ala Lys Thr Lys Arg Tyr Phe Ser Lys Pro Lys Ala Gly Asp 405 410 415 Gln Met Glu Lys Ala Asn Pro Phe His Pro Val Ala Gly Thr Phe Phe 420 425 430 Phe Ser Leu Val Arg Gly Gln Arg Thr Arg Gln Met Leu Tyr Val Lys 435 440 445 Tyr Glu Ile Asp Thr Gln Leu Leu Val Asn Leu Leu Asn Thr Met Ser 450 455 460 Val Leu Val Met Cys Ser Gln Asn Cys Pro Leu Leu Pro Ala Met Thr 465 470 475 480 Arg Glu Ile Phe Asp Leu Cys Ala Phe Val Arg Phe Asn Ala Glu Ala 485 490 495 Arg Val Arg Ala Ala Thr Leu Gln Leu Ile Gly Ile Ala Leu Val Thr 500 505 510 Thr Pro Ala His Val Leu Ala Gln His Phe Ala Glu Ser Leu Asn Glu 515 520 525 Leu Gln Arg Trp Leu Asn Asp Phe Ile Arg Ser Pro Leu Val Gly Gly 530 535 540 Glu Thr Ser Glu Glu Cys Arg Glu Leu Ala 545 550 14 1017 PRT unknown A. thaliana clk-2 putative protein 14 Met Ala Glu Gly Thr Lys Gln Glu Arg Thr Leu Glu Asn Asn Leu Leu 1 5 10 15 His Lys Val Gly Glu Ala Val Ser Ala Ile Ser Asp Ala Lys His Val 20 25 30 Asp Gln Val Ile Ser Ala Ile His Ser Val Ala Val Leu Leu Phe Pro 35 40 45 Val Asp Pro Ser Leu Phe Ser Gly Asn Phe Glu Met Leu His Ile Val 50 55 60 Arg Gly Ser Gly Thr Phe Gly Leu Leu Met Ile Leu Tyr Leu Gly Ser 65 70 75 80 Ala Gln Val Cys Ser Ser Val Val Pro Ser Ala Asp Glu Arg Asn Glu 85 90 95 Trp Leu Glu Thr Phe Tyr Arg Gly Val Ala Phe Pro Thr Phe Ala Arg 100 105 110 Val Leu Leu Leu Asp Val Ala Ser Asp Trp Leu Ser Cys Phe Pro Ile 115 120 125 Ser Val Gln Lys His Leu Tyr Asp Lys Phe Phe Leu Asp Gly Ser Val 130 135 140 Ile Glu Val Val Gln Val Leu Val Pro Phe Leu His His Val Gly Asp 145 150 155 160 Gly Gly Val Asn Ala Asn Ser Val Gln Thr Asn Val Glu Arg Leu Leu 165 170 175 Ile Leu Cys Leu Leu Glu Asn Asp Gly Val Leu Lys Ile Thr Lys Glu 180 185 190 Ile Gly Asn Ile Tyr Gln Gly His Asn Ser Ser Asn Gly Ser Leu Lys 195 200 205 Pro Leu Leu Ser Arg Leu Ser Gln Ile Leu Thr Ser Ile Pro Asp Lys 210 215 220 Ala Arg Ala Ser Cys Thr Glu Ala Asn Cys Thr Val Ile Val Leu Ser 225 230 235 240 Phe Val Gly Glu Val Phe Ser Arg Ile Cys Arg Arg Gly Leu Ser Asp 245 250 255 Leu Leu Leu Ser Glu Val Thr Pro His Val Leu Ala Gln Val Arg Arg 260 265 270 Leu Leu Asn Ser Lys Ile Gly Ala Ile Glu Val Asp Thr Phe Gln Leu 275 280 285 Asp Pro Thr Thr Arg Ile Trp Ser Lys Thr Met Glu Ala Val Thr Asp 290 295 300 Pro Tyr Ala Val Glu Lys Met Ala Glu Gln Leu Leu His Gln Leu Tyr 305 310 315 320 Ala Glu His Pro Ser Asp Val Glu Ala Phe Trp Thr Ile Trp Thr Leu 325 330 335 Phe His Arg Asn Val Ile His Gln Ala Ser Val Arg Gln Ala Lys Cys 340 345 350 Phe Leu Trp Gln Leu Asp Ser Phe Phe Arg Tyr Pro Phe Phe Phe Phe 355 360 365 His Phe His Pro Asn Ala Val Lys Gln Cys Val Leu Glu Cys Pro Pro 370 375 380 Val Thr Asn Thr Leu Ala Lys Gly Asp Val Thr Gln Gly Leu Leu Glu 385 390 395 400 Thr Thr Gln Arg Leu Ala Ser Val Trp Ser Lys Arg Glu Phe Leu Gln 405 410 415 Ser Val Gln Leu Glu Gln Gln Ala Tyr Leu Gln Phe Leu Phe Pro Val 420 425 430 Thr Asp Ile Ser Asp Ile Thr Ala Ala Leu Gly Leu Cys Leu Glu Asn 435 440 445 Met Ser Arg Glu Glu Leu Asp Arg Thr Lys Asp Val Met His Ser Ile 450 455 460 Leu Gln Gly Val Ser Cys Arg Leu Glu Asn Pro Gly Asp Leu Val Arg 465 470 475 480 Lys Met Ala Ser Ser Ile Ala Phe Met Phe Ser Lys Val Ile Asp Pro 485 490 495 Lys Asn Pro Leu Tyr Leu Asp Asp Ser Ile Thr Asp Asn Ala Ile Asp 500 505 510 Trp Glu Phe Gly Leu Gln Thr Ala Ser Ile Thr Asn Thr Met Glu Asn 515 520 525 Gly Asp Gly Glu Asn Lys Arg Ser Ala Ser Leu Thr Glu Val Asn Glu 530 535 540 Ser Ser Arg Arg Asn Lys Gln Lys Glu Asn Arg Lys Ser Lys Asn Ile 545 550 555 560 Ser Ala Phe Val Leu Ala Asp Pro Asn Glu Ile Val Asp Leu Ala Thr 565 570 575 Leu Asn Cys Asp Thr Glu Ser Asp Lys Asp Asp Gly Asp Asp Asp Ala 580 585 590 Ser Val Ser Ser Asp Asn Ser Ser Val Thr Ser Leu Glu Pro Tyr Asp 595 600 605 Leu Met Asp Asp Asp Lys Asp Leu Gly Lys Gln Phe Thr His Leu Val 610 615 620 Asp Val Val Gly Ala Leu Arg Lys Thr Asp Asp Ala Asp Gly Val Glu 625 630 635 640 Lys Ala Ile Tyr Val Ala Glu Lys Leu Val Arg Ala Ser Pro Asp Glu 645 650 655 Leu Thr His Ile Ala Gly Asp Leu Ala Arg Thr Leu Val Gln Val Arg 660 665 670 Cys Ser Asp Ile Ala Ile Glu Gly Glu Glu Asp Ser Ala Glu Glu Lys 675 680 685 Arg Gln Arg Ala Leu Ile Ala Leu Leu Val Thr Arg Pro Phe Glu Ser 690 695 700 Leu Glu Thr Leu Asn Asn Ile Leu Tyr Ser Pro Asn Val Asp Val Ser 705 710 715 720 Gln Arg Ile Met Ile Leu Asp Val Met Ala Glu Ala Ala Arg Glu Leu 725 730 735 Ala Asn Ser Lys Thr Leu Lys Pro Lys His Glu Ala Arg Gly Pro Leu 740 745 750 Ile Ser Asn Ile Ser Asp Pro Gln Pro Trp Tyr Leu Pro Ser Asn Ala 755 760 765 Ser Thr Pro Trp Lys Lys Val Ser Glu Thr Gly Ser Phe His Leu Asn 770 775 780 Trp Ala Asn Arg Phe Glu Arg Glu Leu Gln Ser Lys Pro Gly Gln Thr 785 790 795 800 Lys Lys Gly Lys Ser Arg Arg Trp Ser Leu Lys Ser Ala Asp Arg Asp 805 810 815 Gln Asn Ser Thr Asp Trp Ser Gln Asn Arg Phe Pro Leu Tyr Ala Ala 820 825 830 Ala Phe Met Leu Pro Ala Met Lys Glu Phe Asp Lys Lys Arg His Gly 835 840 845 Val Asp Leu Leu Gly Arg Asp Phe Val Val Leu Gly Lys Leu Val His 850 855 860 Met Leu Gly Val Cys Met Gln Cys Ala Ser Met His Pro Glu Ala Ser 865 870 875 880 Ala Leu Ala Ile Ser Leu Leu Asp Met Leu Gln Arg Arg Glu Val Cys 885 890 895 Asn His Pro Glu Ala Tyr Val Arg Arg Ala Val Leu Phe Ala Ala Ser 900 905 910 Ser Val Leu Val Ser Leu His Pro Ser Tyr Ile Val Ser Thr Leu Val 915 920 925 Glu Gly Asn Leu Asp Leu Ser Arg Ala Leu Glu Trp Ile Arg Thr Trp 930 935 940 Ala Leu Gln Ile Ala Asp Ser Asp Ile Asp Arg Asp Cys Tyr Thr Met 945 950 955 960 Ala Leu Ser Cys Leu Gln Leu His Ala Glu Met Ala Leu Gln Thr Ser 965 970 975 Arg Ala Leu Glu Ser Thr Gly Gly Ser Ser Ser Ser Ser Ser Ile Arg 980 985 990 Pro Met Asn Ile Ser Leu Pro Ser Gly Ile Ser Lys Leu Thr Ser Ile 995 1000 1005 Lys Leu Pro Ser Ser Asn Val His Leu 1010 1015 15 469 DNA Artificial Sequence part of Oryza sativa clk-2 cDNA 15 gtttctactg ggtttggaat ggatacgcac atgggctctt ccatgttgca gaaacagatc 60 ctgatacaga atgcacatca atggctatga ccctccctgc ggctttcatt ctgagatggc 120 ccttcagaca tcacgagcac tggaatcggc ggatcacagc aaggccagca gcagcagcag 180 caggtcgcta ccttctaagc ttgataacat catcatacca tttgccaaca tgatgtgatc 240 gatgtactag attaagttgt aaataagcat atattatctt gccatgtaat attagaaatt 300 ccaggtcagc taattctgat taggcttgat attgtcatat tacagggttt tttaatctgg 360 gatttgaaaa tgctagaatt tttgtcgatt cgtgtggaga ttgtatggaa gttgtcctga 420 tccagattaa agagaattga tgaaaattgt acccaaaaaa aaaaaaaaa 469 16 376 DNA Artificial Sequence part of Oryza sativa clk-2 cDNA 16 agttcctagt gatcagggac ctgctggtgc aggtctntgg agggaagtct cagaatcagg 60 aacacttctg aattggtcac accggtatga aagagaagtt ccatctagat ctggtcaagt 120 taaatcagga aaatctcgta aatggggtct tggaaaagct aaagatttgc agacagagtg 180 gtcaaaaaac agatttcctt tatatgctgc tgcttttatg ctccctgtta tgcaaggata 240 tgataaaaga tcacatggtg ttgacttgct caatcgggac tttgttgtcc taggtaaatt 300 gatatacatg cttggtgtct gtatgaagtg catggcaatg catccagaag catcagctgt 360 tgccccagct cttctt 376 17 122 PRT unknown part of Oryza sativa clk-2 protein 17 Pro Trp Arg Glu Val Ser Glu Ser Gly Thr Leu Leu Asn Trp Ser His 1 5 10 15 Arg Tyr Glu Arg Glu Val Pro Ser Arg Ser Gly Gln Val Lys Ser Gly 20 25 30 Lys Ser Arg Lys Trp Gly Leu Gly Lys Ala Lys Asp Leu Gln Thr Glu 35 40 45 Trp Ser Lys Asn Arg Phe Pro Leu Tyr Ala Ala Ala Phe Met Leu Pro 50 55 60 Val Met Gln Gly Tyr Asp Lys Arg Ser His Gly Val Asp Leu Leu Asn 65 70 75 80 Arg Asp Phe Val Val Leu Gly Lys Leu Ile Tyr Met Leu Gly Val Cys 85 90 95 Met Lys Cys Met Ala Met His Pro Glu Ala Ser Ala Val Ala Pro Ala 100 105 110 Leu Leu Asp Met Ile Arg Ser Arg Ala Val 115 120 18 70 PRT unknown part of Oryza sativa clk-2 protein 18 Leu Glu Trp Ile Arg Thr Trp Ala Leu Pro Cys Cys Arg Asn Tyr Arg 1 5 10 15 Met His Ile Asn Gly Tyr Asp Pro Pro Cys Gly Phe His Ser Glu Met 20 25 30 Ala Leu Gln Thr Ser Arg Ala Leu Glu Ser Ala Asp His Ser Lys Ala 35 40 45 Ser Ser Ser Ser Ser Arg Ser Leu Pro Ser Lys Leu Asp Asn Ile Ile 50 55 60 Ile Pro Phe Ala Asn Met 65 70 19 192 PRT unknown composite protein sequence of Oryza sativa clk-2 protein 19 Pro Trp Arg Glu Val Ser Glu Ser Gly Thr Leu Leu Asn Trp Ser His 1 5 10 15 Arg Tyr Glu Arg Glu Val Pro Ser Arg Ser Gly Gln Val Lys Ser Gly 20 25 30 Lys Ser Arg Lys Trp Gly Leu Gly Lys Ala Lys Asp Leu Gln Thr Glu 35 40 45 Trp Ser Lys Asn Arg Phe Pro Leu Tyr Ala Ala Ala Phe Met Leu Pro 50 55 60 Val Met Gln Gly Tyr Asp Lys Arg Ser His Gly Val Asp Leu Leu Asn 65 70 75 80 Arg Asp Phe Val Val Leu Gly Lys Leu Ile Tyr Met Leu Gly Val Cys 85 90 95 Met Lys Cys Met Ala Met His Pro Glu Ala Ser Ala Val Ala Pro Ala 100 105 110 Leu Leu Asp Met Ile Arg Ser Arg Ala Val Leu Glu Trp Ile Arg Thr 115 120 125 Trp Ala Leu Pro Cys Cys Arg Asn Tyr Arg Met His Ile Asn Gly Tyr 130 135 140 Asp Pro Pro Cys Gly Phe His Ser Glu Met Ala Leu Gln Thr Ser Arg 145 150 155 160 Ala Leu Glu Ser Ala Asp His Ser Lys Ala Ser Ser Ser Ser Ser Arg 165 170 175 Ser Leu Pro Ser Lys Leu Asp Asn Ile Ile Ile Pro Phe Ala Asn Met 180 185 190 20 464 DNA Artificial Sequence part of Glycine max clk-2 cDNA 20 ttttaggaat tggtcaaata gctatgagag ggaacttccc ccaaaaccta atcaggtcaa 60 gaaagggaaa acacgccggt ggagcctaca atctcccaca caacaaaacc agatggagta 120 ttctcataat aagttaccca tgtatgctgc tgcattcatg cttcctgcca tggagggata 180 tgataaaaaa aggcaaggtg ttgacttgct tggaagagat tttattgtct tggggaaact 240 catttatatg cttggggtct gtatgaaatc tgtagccatg catccagaag cttctatgct 300 ggctccttcc ctcctaaata tgttaagatc cagggaggta tgccatcacc aggaagcata 360 tgtgagaaga gccgtgcttt ttgcagctgc atgtgtattg gttgcccttc atcctactta 420 catttcatcc accttactcg aaggaaatgc tgaaatttcg actg 464 21 521 DNA Artificial Sequence part of Glycine max clk-2 cDNA 21 tccggagaag gactctgatt ccccttccaa taaagagaaa agtatttgtt taaagggtaa 60 aaagaagtta ttggacttta atgcgcttga tccagatgag attattgatc cagcatcact 120 gaatcttgaa tcagacgata gcgatgagga tgctgacgat ggtgctagtg agaattcata 180 ttcttcaagt gattcatctt tacggccata tgatttgtca gatgatgact cagatttgaa 240 aagaaaaatt tcacagttgg ctgatgtagt tgcagctctt agaaaatcca atgatgccga 300 tggggtggaa agggctattg atgtagctga aaagctcata agagcatccc ccgatgaact 360 aaaacatgca gcaagggata tgaccagaac tcttgttcag gttcggtgct ctgatatagc 420 tttagaaggt gcagaagaat caactgaaga caaaagacaa agatcattag ttgccttagt 480 agttacctgc ccatttgaat cacttgagtc actaaacaac c 521 22 741 DNA Artificial Sequence part of Glycine max clk-2 cDNA 22 atnnatnnnn nntagcagaa ataattagca gtttagcacg tttagaattt atattctggg 60 ctagaagcta ttcacaatag ctagaagcat cactaatccc cattttagta gggaatttta 120 atggtcactt tggatgcatc agaaggaaga acaggacctg ccttgagtga acttcttact 180 gactccaatg ctcgggaagt ttgaagagcc atctcggcat ggagttgtat acatgtcata 240 gccatcgtat agcattcttt atctgtgtct gactcggcta tgtcaagtgc ccatgtgcga 300 atccattcaa ggccagtcga aatttcagca tttccttcga gtaaggtgga tgaaatgtaa 360 gtaggatgaa gggcaaccaa tacacatgca gctgcaaaaa gcacggctct tctcacatat 420 gcttcctggt gatggcatac ctccctggat cttaacatat ttaggaggga aggagccagc 480 atagaagctt ctggatgcat ggctacagat ttcatacaga ccccaagcat ataaatgagt 540 ttccccaaga caataaaatc tcttccaagc aagtcaacac cttgcctttt tttatcatat 600 ccctccatgg caggaagcat gaatgcagca gcatacatgg gtaacttatt atgagaatac 660 tccatctggt tttgttgtgt gggnnnnngt nnncnccacn nncgtgtttt ccctttcttg 720 acctgattng nttttgnnng a 741 23 456 DNA Artificial Sequence part of Glycine max clk-2 cDNA 23 ttgaaatttc aactggcctt gaatggattc gcacatgggc acttgatgta gccgagtcgg 60 acacagataa agaatgctat acgatggcta tgacatgtat acagctccat gttgagatgg 120 ctcttcaaac ttcccgagca ttggagtcag taagaaattc actcaaggca ggtcctgttc 180 ttccttctga tgcatccaaa gtgaccatta aaattcccca cttaaatggg gattagtgat 240 gcttctagct attgtggatg gattctagcc cagaatacaa attctatatc atgctaaact 300 gctaattatt tctgctactc tattaattta atgttttatg gggtccttgt tttcgtaaga 360 aattgtatat tttaggattg tatatattct cacaagggga cgaggttgat gtcaatgctc 420 ccctacacac ccacccttga tgtagcacgt gttact 456 24 455 DNA Artificial Sequence part of Glycine max clk-2 cDNA 24 aaactcgcgg cggaaggcag cagagtggcg ccgatggagg agggtttaga gaagagagaa 60 ttggaaggcg aagtagttac caaggttgtc gaagtggttt cggctataaa gaatgcgaag 120 cacgtcgatc aagtcattcg cgcgcttcat tccttagtca cccttctttt cccctttgac 180 tcttcactcc tatcagatag cattgaccag agttaccgag accaggtcga agttccttct 240 gcagaaaaac gacatgcttg gtggcgtgcg ttttatcgag gagctgcttt tcctacactg 300 gctaggtttt tattacttga tgttgcctcg aactggttgg gttgttttcc ctttatggcg 360 cagaagtaca tttatgatgt tttctttgtt cgtggattgg tcactgacgt tctgcagatt 420 ctggttcctt ttcttcagct gagtgcgagt gatgg 455 25 208 PRT unknown part of Glycine max clk-2 protein 25 Lys Xaa Asn Gln Val Lys Lys Gly Lys Thr Arg Xaa Trp Xaa Xaa Xaa 1 5 10 15 Xaa Pro Thr Gln Gln Asn Gln Met Glu Tyr Ser His Asn Lys Leu Pro 20 25 30 Met Tyr Ala Ala Ala Phe Met Leu Pro Ala Met Glu Gly Tyr Asp Lys 35 40 45 Lys Arg Gln Gly Val Asp Leu Leu Gly Arg Asp Phe Ile Val Leu Gly 50 55 60 Lys Leu Ile Tyr Met Leu Gly Val Cys Met Lys Ser Val Ala Met His 65 70 75 80 Pro Glu Ala Ser Met Leu Ala Pro Ser Leu Leu Asn Met Leu Arg Ser 85 90 95 Arg Glu Val Cys His His Gln Glu Ala Tyr Val Arg Arg Ala Val Leu 100 105 110 Phe Ala Ala Ala Cys Val Leu Val Ala Leu His Pro Thr Tyr Ile Ser 115 120 125 Ser Thr Leu Leu Glu Gly Asn Ala Glu Ile Ser Thr Gly Leu Glu Trp 130 135 140 Ile Arg Thr Trp Ala Leu Asp Ile Ala Glu Ser Asp Thr Asp Lys Glu 145 150 155 160 Cys Tyr Thr Met Ala Met Thr Cys Ile Gln Leu His Ala Glu Met Ala 165 170 175 Leu Gln Thr Ser Arg Ala Leu Glu Ser Val Arg Ser Ser Leu Lys Ala 180 185 190 Gly Pro Val Leu Pro Ser Asp Ala Ser Lys Val Thr Ile Lys Ile Pro 195 200 205 26 151 PRT unknown part of Glycine max clk-2 protein 26 Asn Trp Ser Asn Ser Tyr Glu Arg Glu Leu Pro Pro Lys Pro Asn Gln 1 5 10 15 Val Lys Lys Gly Lys Thr Arg Arg Trp Ser Leu Gln Ser Pro Thr Gln 20 25 30 Gln Asn Gln Met Glu Tyr Ser His Asn Lys Leu Pro Met Tyr Ala Ala 35 40 45 Ala Phe Met Leu Pro Ala Met Glu Gly Tyr Asp Lys Lys Arg Gln Gly 50 55 60 Val Asp Leu Leu Gly Arg Asp Phe Ile Val Leu Gly Lys Leu Ile Tyr 65 70 75 80 Met Leu Gly Val Cys Met Lys Ser Val Ala Met His Pro Glu Ala Ser 85 90 95 Met Leu Ala Pro Ser Leu Leu Asn Met Leu Arg Ser Arg Glu Val Cys 100 105 110 His His Gln Glu Ala Tyr Val Arg Arg Ala Val Leu Phe Ala Ala Ala 115 120 125 Cys Val Leu Val Ala Leu His Pro Thr Tyr Ile Ser Ser Thr Leu Leu 130 135 140 Glu Gly Asn Ala Glu Ile Ser 145 150 27 172 PRT unknown part of Glycine max clk-2 protein 27 Glu Lys Asp Ser Asp Ser Pro Ser Asn Lys Glu Lys Ser Ile Cys Leu 1 5 10 15 Lys Gly Lys Lys Lys Leu Leu Asp Phe Asn Ala Leu Asp Pro Asp Glu 20 25 30 Ile Ile Asp Pro Ala Ser Leu Asn Leu Glu Ser Asp Asp Ser Asp Glu 35 40 45 Asp Ala Asp Asp Gly Ala Ser Glu Asn Ser Tyr Ser Ser Ser Asp Ser 50 55 60 Ser Leu Arg Pro Tyr Asp Leu Ser Asp Asp Asp Ser Asp Leu Lys Arg 65 70 75 80 Lys Ile Ser Gln Leu Ala Asp Val Val Ala Ala Leu Arg Lys Ser Asn 85 90 95 Asp Ala Asp Gly Val Glu Arg Ala Ile Asp Val Ala Glu Lys Leu Ile 100 105 110 Arg Ala Ser Pro Asp Glu Leu Lys His Ala Ala Arg Asp Met Thr Arg 115 120 125 Thr Leu Val Gln Val Arg Cys Ser Asp Ile Ala Leu Glu Gly Ala Glu 130 135 140 Glu Ser Thr Glu Asp Lys Arg Gln Arg Ser Leu Val Ala Leu Val Val 145 150 155 160 Thr Cys Pro Phe Glu Ser Leu Glu Ser Leu Asn Asn 165 170 28 134 PRT unknown part of Glycine max clk-2 protein 28 Met Glu Glu Gly Leu Glu Lys Arg Glu Leu Glu Gly Glu Val Val Thr 1 5 10 15 Lys Val Val Glu Val Val Ser Ala Ile Lys Asn Ala Lys His Val Asp 20 25 30 Gln Val Ile Arg Ala Leu His Ser Leu Val Thr Leu Leu Phe Pro Phe 35 40 45 Asp Ser Ser Leu Leu Ser Asp Ser Ile Asp Gln Ser Tyr Arg Asp Gln 50 55 60 Val Glu Val Pro Ser Ala Glu Lys Arg His Ala Trp Trp Arg Ala Phe 65 70 75 80 Tyr Arg Gly Ala Ala Phe Pro Thr Leu Ala Arg Phe Leu Leu Leu Asp 85 90 95 Val Ala Ser Asn Trp Leu Gly Cys Phe Pro Phe Met Ala Gln Lys Tyr 100 105 110 Ile Tyr Asp Val Phe Phe Val Arg Gly Leu Val Thr Asp Val Leu Gln 115 120 125 Ile Leu Val Pro Phe Leu 130 29 75 PRT unknown part of Glycine max clk-2 protein 29 Glu Ile Ser Thr Gly Leu Glu Trp Ile Arg Thr Trp Ala Leu Asp Val 1 5 10 15 Ala Glu Ser Asp Thr Asp Lys Glu Cys Tyr Thr Met Ala Met Thr Cys 20 25 30 Ile Gln Leu His Val Glu Met Ala Leu Gln Thr Ser Arg Ala Leu Glu 35 40 45 Ser Val Arg Asn Ser Leu Lys Ala Gly Pro Val Leu Pro Ser Asp Ala 50 55 60 Ser Lys Val Thr Ile Lys Ile Pro His Leu Asn 65 70 75 30 528 PRT unknown composite of Glycine max clk-2 protein 30 Met Glu Glu Gly Leu Glu Lys Arg Glu Leu Glu Gly Glu Val Val Thr 1 5 10 15 Lys Val Val Glu Val Val Ser Ala Ile Lys Asn Ala Lys His Val Asp 20 25 30 Gln Val Ile Arg Ala Leu His Ser Leu Val Thr Leu Leu Phe Pro Phe 35 40 45 Asp Ser Ser Leu Leu Ser Asp Ser Ile Asp Gln Ser Tyr Arg Asp Gln 50 55 60 Val Glu Val Pro Ser Ala Glu Lys Arg His Ala Trp Trp Arg Ala Phe 65 70 75 80 Tyr Arg Gly Ala Ala Phe Pro Thr Leu Ala Arg Phe Leu Leu Leu Asp 85 90 95 Val Ala Ser Asn Trp Leu Gly Cys Phe Pro Phe Met Ala Gln Lys Tyr 100 105 110 Ile Tyr Asp Val Phe Phe Val Arg Gly Leu Val Thr Asp Val Leu Gln 115 120 125 Ile Leu Val Pro Phe Leu Glu Lys Asp Ser Asp Ser Pro Ser Asn Lys 130 135 140 Glu Lys Ser Ile Cys Leu Lys Gly Lys Lys Lys Leu Leu Asp Phe Asn 145 150 155 160 Ala Leu Asp Pro Asp Glu Ile Ile Asp Pro Ala Ser Leu Asn Leu Glu 165 170 175 Ser Asp Asp Ser Asp Glu Asp Ala Asp Asp Gly Ala Ser Glu Asn Ser 180 185 190 Tyr Ser Ser Ser Asp Ser Ser Leu Arg Pro Tyr Asp Leu Ser Asp Asp 195 200 205 Asp Ser Asp Leu Lys Arg Lys Ile Ser Gln Leu Ala Asp Val Val Ala 210 215 220 Ala Leu Arg Lys Ser Asn Asp Ala Asp Gly Val Glu Arg Ala Ile Asp 225 230 235 240 Val Ala Glu Lys Leu Ile Arg Ala Ser Pro Asp Glu Leu Lys His Ala 245 250 255 Ala Arg Asp Met Thr Arg Thr Leu Val Gln Val Arg Cys Ser Asp Ile 260 265 270 Ala Leu Glu Gly Ala Glu Glu Ser Thr Glu Asp Lys Arg Gln Arg Ser 275 280 285 Leu Val Ala Leu Val Val Thr Cys Pro Phe Glu Ser Leu Glu Ser Leu 290 295 300 Asn Asn Glu Ile Ser Thr Gly Leu Glu Trp Ile Arg Thr Trp Ala Leu 305 310 315 320 Asp Val Ala Glu Ser Asp Thr Asp Lys Glu Cys Tyr Thr Met Ala Met 325 330 335 Thr Cys Ile Gln Leu His Val Glu Met Ala Leu Gln Thr Ser Arg Ala 340 345 350 Leu Glu Ser Val Arg Asn Ser Leu Lys Ala Gly Pro Val Leu Pro Ser 355 360 365 Asp Ala Ser Lys Val Thr Ile Lys Ile Pro His Leu Asn Asn Trp Ser 370 375 380 Asn Ser Tyr Glu Arg Glu Leu Pro Pro Lys Pro Asn Gln Val Lys Lys 385 390 395 400 Gly Lys Thr Arg Arg Trp Ser Leu Gln Ser Pro Thr Gln Gln Asn Gln 405 410 415 Met Glu Tyr Ser His Asn Lys Leu Pro Met Tyr Ala Ala Ala Phe Met 420 425 430 Leu Pro Ala Met Glu Gly Tyr Asp Lys Lys Arg Gln Gly Val Asp Leu 435 440 445 Leu Gly Arg Asp Phe Ile Val Leu Gly Lys Leu Ile Tyr Met Leu Gly 450 455 460 Val Cys Met Lys Ser Val Ala Met His Pro Glu Ala Ser Met Leu Ala 465 470 475 480 Pro Ser Leu Leu Asn Met Leu Arg Ser Arg Glu Val Cys His His Gln 485 490 495 Glu Ala Tyr Val Arg Arg Ala Val Leu Phe Ala Ala Ala Cys Val Leu 500 505 510 Val Ala Leu His Pro Thr Tyr Ile Ser Ser Thr Leu Leu Glu Gly Asn 515 520 525 31 876 PRT unknown C. elegans clk-2 (QM37) mutant protein, with C to Y substitution at position 772 31 Met Asn Leu Arg Ser Arg Leu Val Asn Ala Thr Glu Arg Ala Val Leu 1 5 10 15 Phe Gln Ile Phe Lys Asp Val Gln Asn Asp Pro Glu Lys Tyr Asp Asn 20 25 30 Ala Val Glu Ala Ile Cys Glu Ser Ile Asp Tyr Phe Gly Lys Phe Leu 35 40 45 Thr Asp Ser Glu Tyr Leu Thr Gln Ile Lys Pro Ile Leu Asp Thr Gln 50 55 60 Cys Pro Thr Lys Ser Ile Ile Cys Phe Ser Lys Cys Leu Thr Lys Val 65 70 75 80 Ser Thr Asp Ile Asn Thr Thr Thr Phe Arg Asp Val Ile Thr Met Leu 85 90 95 Asp Trp Leu Lys Tyr Val Val Glu Lys Ser Leu Thr Ser Ala Ile Cys 100 105 110 Ser Ser Leu Lys Val Lys Glu Thr Asp Val Ser Ala Val Gln Leu Tyr 115 120 125 Arg Glu Phe Ala Ser Ala Cys Ser Asn Ile Pro Glu Lys Val Ser Asn 130 135 140 Cys Cys Ala Lys Ala Leu Ser Gly Glu His Val Lys Tyr Ile Asn Thr 145 150 155 160 Val Lys Trp Ile Phe Lys Met Asn Leu Val Gln Gly Ile Gln Lys Ala 165 170 175 Met Leu Leu Ala His Asp Asp Ile Val Thr Ala Ala Pro Phe Thr Ser 180 185 190 Phe Tyr Gly Ser Gly Gly Pro Tyr Met Lys Thr Val Ala Glu Ile Ile 195 200 205 Ser Ser Gly Arg Ile Asp Ile Thr Asn Lys Asp Gly Leu Leu Val Gln 210 215 220 Met Ile Glu Trp Ile Gly Ser Leu Asn Asn Phe Asp Ser Gln Trp Arg 225 230 235 240 Arg Met Met Phe Leu Ile Phe Gln Glu Pro Thr Tyr Gln Gly Ile Gln 245 250 255 Val His Glu Ser Leu Leu Thr Thr Leu Phe Leu Ile Ser Lys Ser Asp 260 265 270 Gln Ile Leu Lys Arg Cys Ile Glu Ala Thr Asp Leu Thr Gly Thr Leu 275 280 285 Lys Arg Val Val Met Val Lys Leu Pro Phe Gln Arg Val Leu Lys Arg 290 295 300 Lys Thr Ile Glu Ile Leu Ile Asn Phe Val Tyr Arg Thr Lys Glu Gln 305 310 315 320 Phe Ala Ile Gln Leu Leu Glu Thr Ser Val Lys Ile Trp Ser Asp Leu 325 330 335 Asn Tyr Ala Lys Ser Ala Pro Glu Ser Gln Glu Arg His Ile Val Arg 340 345 350 Met Ile Leu Tyr Leu Val His Leu Phe Arg Thr Cys Ser Ser Ile Asp 355 360 365 Trp Glu Ser Leu Phe Leu Asn Ser Met Asp Gly Val His Cys Arg Met 370 375 380 Ser Met Leu Pro Met Tyr Val Gln Ser Gly Ile Phe Val Asn Gln Ala 385 390 395 400 Leu Cys Lys Gln Ala Thr Lys His Arg Ser Lys Thr His Gly Ser Asp 405 410 415 Glu Gln Pro Pro Glu Thr Leu Glu Glu Asn Lys Phe Val Ser Ser Glu 420 425 430 Val Gly Lys Ile Trp Phe Glu Glu Met Thr Ser Ile Leu Glu His Gly 435 440 445 Phe Asn Ser Ser Thr Val Lys Asp Ser Glu Arg Val Arg Glu Thr Ala 450 455 460 Asn Glu Ile Thr Lys Asp Asp Ser Gly Glu Glu Phe Glu Glu Thr Asn 465 470 475 480 Ala Gln Arg Leu Gln Asn Asn Lys Asp Ser Ala Ala Ile Thr Ser Lys 485 490 495 Asn Asn Leu Arg Leu Asp Ser Asp Asp Asp Glu Asp Phe Pro Asp Tyr 500 505 510 Gln Val Asn Glu Ser Glu Lys Ile Phe Lys Asn Leu Glu Ile Gly Glu 515 520 525 Glu Pro Lys Asn Lys Val Thr Pro Pro Ala Tyr Ile Ala Asp Ala Phe 530 535 540 Glu Met Leu Leu Glu Lys Glu Lys Tyr Glu Val Phe Glu Ala Ala Phe 545 550 555 560 Phe Asn Ile Thr Asn Leu Ile Asn Arg Arg Pro Ile Gly Phe Pro Gln 565 570 575 Ile Ala Glu Lys Leu Phe Ile Arg Ile Leu His Leu Gln Asn Asn Phe 580 585 590 Gly Thr Pro Lys Phe Lys Glu Thr Val Asp Glu Ile Ala Val Ala Cys 595 600 605 Ile Thr Gln Arg Pro Glu Ile Val Pro Ser Val Val Arg Leu Ile Ile 610 615 620 Ala Pro Gly Gln Gly Phe Ser Ile Lys Gln Arg Leu Leu His Tyr Ile 625 630 635 640 His Asn Ala Ala Asp Gly Met Gly Ala Leu Asp Lys Lys Leu Glu Glu 645 650 655 Cys Val Met Ala Gln Gln Leu Arg Ile Gly Gly Pro Thr Leu Ser Ile 660 665 670 Ile Leu His Arg Thr Ile Asn Thr Asp Tyr Asp Asp Glu Asp Glu Asp 675 680 685 Pro His Arg Leu Leu Val Pro Glu Trp Arg Arg Met Val Asp Ala Arg 690 695 700 Ile Ala Ala Asn Thr Arg Arg Ile Gly Thr Thr Arg Glu Pro Pro Arg 705 710 715 720 Ala Gly Val Val Asn Arg Leu Ala Gln Ala Ala Lys Tyr Met Phe Tyr 725 730 735 Pro Leu Leu Val Leu Pro Arg Gly Glu Asn Ala Ser Leu Leu Gly Lys 740 745 750 Asp Ser Asp Leu Leu Ala Ser Leu Ile Met Val Ala Ser Met Val Tyr 755 760 765 Val Arg Tyr Gly Val Cys Pro Gln Ile His Arg Met Ser Ser Glu Leu 770 775 780 Ile Ser Tyr Ala Thr Pro His Arg Phe Ser Glu Asn Ala Lys Leu Arg 785 790 795 800 Thr Ala Cys Ile Ile Ala His Leu Asn Val Thr Thr Leu Leu Pro Gly 805 810 815 Asp Leu Met Asp Glu Leu Phe Asp Val Pro Ala Leu Ile Gly Trp Phe 820 825 830 Asp Trp Ala Asn Ser Val Leu Val Asn Ala Ser Ser Ser Gln Leu Glu 835 840 845 Lys Asp Met Thr Arg Gln Phe Gly His Ser Val Thr Lys His Leu Gln 850 855 860 Arg His His Pro Ala Val Leu Gln His Gln Asp Val 865 870 875 32 688 PRT unknown Tel2p, S. cerevisiae clk-2 protein 32 Met Val Leu Glu Thr Leu Lys Gln Gly Leu Asp Ser Ser Gln Ile His 1 5 10 15 Glu Ala Leu Ile Gln Leu Asp Ser Tyr Pro Arg Glu Pro Val Asp Leu 20 25 30 Asp Ala Ser Met Val Leu Ile Lys Phe Val Ile Pro Val Tyr Pro Ser 35 40 45 Leu Pro Glu Arg Ser Lys Val Ile Leu Arg Arg Leu Ala Ser Lys Ser 50 55 60 Phe Thr Phe Leu Cys Gln Ile Val Thr Phe Ser Arg Thr Ile Ser Gly 65 70 75 80 Arg Asp Gly Leu Gln Glu Ile Arg Ile Tyr Gln Glu Ile Leu Glu Asp 85 90 95 Ile Ile Ser Phe Glu Pro Gly Cys Leu Thr Phe Tyr Leu Lys Ala Ser 100 105 110 Thr Thr Ser Lys Ala Asp Arg Asp Ser Ile Lys Ala Leu Phe Phe Gly 115 120 125 Ser Lys Leu Phe Asn Val Leu Ala Asn Arg Ile Asp Met Ala Lys Tyr 130 135 140 Leu Gly Tyr Leu Arg Leu Gln Trp Lys Phe Leu Leu Glu Ser Asn Glu 145 150 155 160 Thr Asp Pro Pro Gly Phe Leu Gly Glu Trp Leu Val Ser Ser Phe Leu 165 170 175 Leu Asn Pro Val Leu Ala Ala Asp Met Leu Leu Gly Glu Leu Phe Leu 180 185 190 Leu Lys Glu Ser Tyr Phe Phe Ser Phe Gln Lys Ile Ile Ser Ala Ser 195 200 205 Ser Leu Ile Asp Gln Lys Arg Leu Ile Ala Lys Phe Leu Leu Pro Tyr 210 215 220 Ile Gln Val Ile Val Thr Leu Glu Asn Leu Asn Asp Val Arg Lys Ile 225 230 235 240 Leu Arg Arg Phe Asp Leu Asp Lys Ile Ile Ser Leu Ser Val Leu Phe 245 250 255 Glu Ile Gln Ser Leu Pro Leu Lys Glu Val Ile Val Arg Leu Met Ser 260 265 270 Asn His Ser Ser Thr Lys Phe Val Ser Ala Leu Val Ser Lys Phe Ala 275 280 285 Asp Phe Thr Asp Glu Glu Val Asp Thr Lys Thr Cys Glu Leu Leu Val 290 295 300 Leu Phe Ala Val His Asn Leu Asn His Ser Gln Arg Glu Glu Ile Ala 305 310 315 320 His Asp Glu Arg Phe Leu Asn Gly Val Thr Lys His Leu Gly Ser Asn 325 330 335 Glu Arg Glu Ala Arg Glu Arg Ala Met Phe Ile Ala Lys Leu Leu Ser 340 345 350 Gly Gly His Leu Lys Tyr Glu Ser Asp Phe Lys Ile Asn Ile Pro Asn 355 360 365 Val Lys Phe Glu Ser Asn Ser Asp Asp Lys Ile Ile Asp Phe Gln Ser 370 375 380 Leu Lys Asn Pro Ser Ile Cys Asn Thr Gln Thr Asp Val Gly Lys Asp 385 390 395 400 Lys Ile Thr Glu Val Ser Gly His Val Gln Ser Leu Thr Leu Asp Cys 405 410 415 Ser Asp Ser Asp Asp Glu Asp Glu Asn Asp Glu Arg Glu Ile Val Lys 420 425 430 Arg Ile Val Phe Leu Lys Asp Leu Met Lys Glu Tyr Glu Lys Thr Gly 435 440 445 Glu Ser Arg Lys Ala Pro Leu Ile Pro Leu Leu Lys Gln Thr Val Lys 450 455 460 Leu Ile Arg Gln Lys Ala Asp Phe Gln Leu Glu Val Gly Tyr Tyr Ala 465 470 475 480 Gln Gly Ile Leu Ser Ser Ile Val Cys Leu Asn Asn Glu Phe Asp Glu 485 490 495 Pro Leu Phe Glu Gln Trp Arg Ile Asn Ala Leu Thr Ser Ile Leu Val 500 505 510 Val Leu Pro Glu Lys Val Asn Gly Ala Ile Asn Ile Leu Phe Asn Ser 515 520 525 Glu Leu Ser Leu Gln Gln Arg Met Ser Leu Leu Ser Ala Leu Gly Leu 530 535 540 Ser Ala Arg Glu Leu Arg Gly Leu Asp Asp Pro Thr Ile Val Lys Pro 545 550 555 560 Lys Phe Asp Phe Pro Thr Asn Arg Leu Pro Trp Asp Asp Gln Ser His 565 570 575 His Asn Ser Arg Leu Val Glu Val Gln Glu Ser Thr Ser Met Ile Lys 580 585 590 Lys Thr Lys Thr Val Trp Lys Ser Arg Lys Leu Gly Lys Asp Arg Glu 595 600 605 Lys Gly Thr Gln Asn Arg Phe Arg Lys Tyr Ala Gly Leu Phe Phe Tyr 610 615 620 Pro Leu Ala His Gly Trp Leu Asn Gly Ile Asp Val Gly Thr Tyr Asn 625 630 635 640 Gln Leu Phe Lys Ser His Tyr Leu Thr Thr Leu Arg Ile Ile Tyr Ser 645 650 655 Cys Ala Asn Pro Val His Asp Phe Glu Ser Met Thr Glu Leu Met Asn 660 665 670 His Ile Ile Ser Ser Ala Ile Glu Glu Gly Ile Ser Leu Asn Lys Gly 675 680 685 33 1520 DNA C. elegans cex-7 cDNA 33 tcgaacttca atttatatcg atgtttcttc tattttagtg caaattttaa tcaaaaaaat 60 taaacctttt gtcaacatgg cgcgaaattt tcccactgct ctcgattacc tgggatcaga 120 agctgaagat tttaacaagg cgcaacactt gtatctgaaa ccgatggctg ttataaaaat 180 taccgtagtg ctgcctcgga tgacgatccc gggacaatca atatctaatt gggatctcat 240 ggaaagacta aaacgtgcaa ttgatccaat tcaaatggat tcttgcaaag ttcgtgagag 300 caatatcgac agtgttattt ttgaagcgga acttctttcg ctaggaatca tgcagaaaac 360 gatgaagatt ctcgatggat tctctatgaa agtgtctgga tttgctgagc ccttgaaagt 420 taaaacaaaa gaggcaaagc ttgattttcc aagtcgtcat gattgggatg attggtttat 480 gaaacataag atggacgaga tgaaacccgg agaacgtcct gatacagttt atttggcaag 540 aattccagtg aaatggttct gcgatggtta caatgatctt ccttctgaac gacgtcttcg 600 cgttgcaatg gaagcgttcg gatctgttag agttgtcgac attccaattt gtgacccact 660 ccgatctcga atgaattcaa aaatctctgg tattcagcaa aaaggtttcg gattagggca 720 ggatgtattt tttgaagcat atgtgcaatt tatggagtat aaaggtttcg ccactgccat 780 ggattccctg agaaatcgta aatgggcaaa acgaattgat ggacggttct ttcaggctaa 840 tgtcaaggtg gatttcgatc gatcacgtca tctcagcgaa gttcaaattg caaagcgagc 900 ggaagaacga cgtcaaattg aaacggaacg actgcggcaa gaagaagagg agttgaatat 960 cagacgtcaa gaagaactga aagttaaaca agaactcgat gataaagata gaagacgaga 1020 agatagagaa cggaaacgcc gcgaaaagag ggaactcgaa cgaatggctg aagaagagaa 1080 aaaacgcctt gaaaaggaac gacttgaagc agagcaacga tcacgagcca ctcgccgttt 1140 gcaaggtgtt cgtcttttga agtttctgtt tgaaaaaatt gaagcacgag aagagagacg 1200 aaagagaaaa gaagaagaaa agttgaaaga tgagctgagc aaaatcaaag aacttgaaga 1260 acaacctgta gaacaagaag acgcattgag gcaagctcta cttcagcaga gggaaattcg 1320 aatgcgagaa cgcttgaaag agaagatgaa agcttcagga gcagagaagg ataagaagag 1380 agataaaaat aagacttcta gaagaagacg aactaataga catgacactt catcctcttc 1440 atccgaagat tctgattctc catctgattc tccccactct cgtcgacaaa gaaaacgtca 1500 aagtgaagga gatgatttcc 1520 34 478 PRT unknown C. elegans cex-7 protein 34 Met Ala Arg Asn Phe Pro Leu Tyr Leu Gly Ser Glu Ala Glu Asp Phe 1 5 10 15 Asn Lys Ala Gln His Leu Tyr Leu Lys Pro Met Ala Val Ile Lys Ile 20 25 30 Thr Val Val Leu Pro Arg Met Thr Ile Pro Gly Gln Ser Ile Ser Asn 35 40 45 Trp Asp Leu Met Glu Arg Leu Lys Arg Ala Ile Asp Pro Ile Gln Met 50 55 60 Asp Ser Cys Lys Val Arg Glu Ser Asn Ile Asp Ser Val Ile Phe Glu 65 70 75 80 Ala Glu Leu Leu Ser Leu Gly Ile Met Gln Lys Thr Met Lys Ile Leu 85 90 95 Asp Gly Phe Ser Met Lys Val Ser Gly Phe Ala Glu Pro Leu Lys Val 100 105 110 Lys Thr Lys Glu Ala Lys Leu Asp Phe Pro Ser Arg His Asp Trp Asp 115 120 125 Asp Trp Phe Met Lys His Lys Met Asp Glu Met Lys Pro Gly Glu Arg 130 135 140 Pro Asp Thr Val Tyr Leu Ala Arg Ile Pro Val Lys Trp Phe Cys Asp 145 150 155 160 Gly Tyr Asn Asp Leu Pro Ser Glu Arg Arg Leu Arg Val Ala Met Glu 165 170 175 Ala Phe Gly Ser Val Arg Val Val Asp Ile Pro Ile Cys Asp Pro Leu 180 185 190 Arg Ser Arg Met Asn Ser Lys Ile Ser Gly Ile Gln Gln Lys Gly Phe 195 200 205 Gly Leu Gly Gln Asp Val Phe Phe Glu Ala Tyr Val Gln Phe Met Glu 210 215 220 Tyr Lys Gly Phe Ala Thr Ala Met Asp Ser Leu Arg Asn Arg Lys Trp 225 230 235 240 Ala Lys Arg Ile Asp Gly Arg Phe Phe Gln Ala Asn Val Lys Val Asp 245 250 255 Phe Asp Arg Ser Arg His Leu Ser Glu Val Gln Ile Ala Lys Arg Ala 260 265 270 Glu Glu Arg Arg Gln Ile Glu Thr Glu Arg Leu Arg Gln Glu Glu Glu 275 280 285 Glu Leu Asn Ile Arg Arg Gln Glu Glu Leu Lys Val Lys Gln Glu Leu 290 295 300 Asp Asp Lys Asp Arg Arg Arg Glu Asp Arg Glu Arg Lys Arg Arg Glu 305 310 315 320 Lys Arg Glu Leu Glu Arg Met Ala Glu Glu Glu Lys Lys Arg Leu Glu 325 330 335 Lys Glu Arg Leu Glu Ala Glu Gln Arg Ser Arg Ala Thr Arg Arg Leu 340 345 350 Gln Gly Val Arg Leu Leu Lys Phe Leu Phe Glu Lys Ile Glu Ala Arg 355 360 365 Glu Glu Arg Arg Lys Arg Lys Glu Glu Glu Lys Leu Lys Asp Glu Leu 370 375 380 Ser Lys Ile Lys Glu Leu Glu Glu Gln Pro Val Glu Gln Glu Asp Ala 385 390 395 400 Leu Arg Gln Ala Leu Leu Gln Gln Arg Glu Ile Arg Met Arg Glu Arg 405 410 415 Leu Lys Glu Lys Met Lys Ala Ser Gly Ala Glu Lys Asp Lys Lys Arg 420 425 430 Asp Lys Asn Lys Thr Ser Arg Arg Arg Arg Thr Asn Arg His Asp Thr 435 440 445 Ser Ser Ser Ser Ser Glu Asp Ser Asp Ser Pro Ser Asp Ser Pro His 450 455 460 Ser Arg Arg Gln Arg Lys Arg Gln Ser Glu Gly Asp Asp Phe 465 470 475 35 550 PRT unknown XE7, Homo sapiens cex-7 protein 35 Met Ala Ala Ala Thr Ile Val His Asp Thr Ser Glu Ala Val Glu Leu 1 5 10 15 Cys Pro Ala Tyr Gly Leu Tyr Leu Lys Pro Ile Thr Lys Met Thr Ile 20 25 30 Ser Val Ala Leu Pro Gln Leu Lys Gln Pro Gly Lys Ser Ile Ser Asn 35 40 45 Trp Glu Val Met Glu Arg Leu Lys Gly Met Val Gln Asn His Gln Phe 50 55 60 Ser Thr Leu Arg Ile Ser Lys Ser Thr Met Asp Phe Ile Arg Phe Glu 65 70 75 80 Gly Glu Val Glu Asn Lys Ser Leu Val Lys Ser Phe Leu Ala Cys Leu 85 90 95 Asp Gly Lys Thr Ile Lys Leu Ser Gly Phe Ser Asp Ile Leu Lys Val 100 105 110 Arg Ala Ala Glu Phe Lys Ile Asp Phe Pro Thr Arg His Asp Trp Asp 115 120 125 Ser Phe Phe Arg Asp Ala Lys Asp Met Asn Glu Thr Leu Pro Gly Glu 130 135 140 Arg Pro Asp Thr Ile His Leu Glu Gly Leu Pro Cys Lys Trp Phe Ala 145 150 155 160 Leu Lys Glu Ser Gly Ser Glu Lys Pro Ser Glu Asp Val Leu Val Lys 165 170 175 Val Phe Glu Lys Phe Gly Glu Ile Arg Asn Val Asp Ile Pro Met Leu 180 185 190 Asp Pro Tyr Arg Glu Glu Met Thr Gly Arg Asn Phe His Thr Phe Ser 195 200 205 Phe Gly Gly His Leu Asn Phe Glu Ala Tyr Val Gln Tyr Arg Glu Tyr 210 215 220 Met Gly Phe Ile Gln Ala Met Ser Ala Leu Arg Gly Met Lys Leu Met 225 230 235 240 Tyr Lys Gly Glu Asp Gly Lys Ala Val Ala Cys Asn Ile Lys Val Ser 245 250 255 Phe Asp Ser Thr Lys His Leu Ser Asp Ala Ser Ile Lys Lys Arg Gln 260 265 270 Leu Glu Arg Gln Lys Leu Gln Glu Leu Glu Gln Gln Arg Glu Glu Gln 275 280 285 Lys Arg Arg Glu Lys Glu Ala Glu Glu Arg Gln Arg Ala Glu Glu Arg 290 295 300 Lys Gln Lys Glu Leu Glu Glu Leu Glu Arg Glu Arg Lys Arg Glu Glu 305 310 315 320 Lys Leu Arg Lys Arg Glu Gln Lys Gln Arg Asp Arg Glu Leu Arg Arg 325 330 335 Asn Gln Lys Lys Leu Glu Lys Leu Gln Ala Glu Glu Gln Lys Gln Leu 340 345 350 Gln Glu Lys Ile Lys Leu Glu Glu Arg Lys Leu Leu Leu Ala Gln Arg 355 360 365 Asn Leu Gln Ser Ile Arg Leu Ile Ala Glu Leu Leu Ser Arg Ala Lys 370 375 380 Ala Val Lys Leu Arg Glu Gln Glu Gln Lys Glu Glu Lys Leu Arg Leu 385 390 395 400 Gln Gln Gln Glu Glu Arg Arg Arg Leu Gln Glu Ala Glu Leu Arg Arg 405 410 415 Val Glu Glu Glu Lys Glu Arg Ala Leu Gly Leu Gln Arg Lys Glu Arg 420 425 430 Glu Leu Arg Glu Arg Leu Leu Ser Ile Leu Gln Ser Lys Lys Pro Asp 435 440 445 Asp Ser His Thr His Asp Glu Leu Gly Val Ala His Gly Pro Ala Ala 450 455 460 Ala Arg Pro Gly His Pro Ala Asp Arg Val Val Arg Leu Cys Glu Arg 465 470 475 480 His His Ala Ala Pro Pro Arg Gly Pro Ala Pro Gly Arg Cys Pro Gln 485 490 495 Gly Glu Pro Gly Pro Pro Arg Gly Arg Arg Arg Ser Gln Lys Arg Glu 500 505 510 Arg Glu Arg Gly Arg Gly Gly Pro Met Gln Gly Gly Ser Glu Leu Leu 515 520 525 Ser Cys Gly Pro Arg Gly Trp Leu Ser Arg Glu Glu Val Pro Gly Arg 530 535 540 Arg Pro Leu Leu His Ser 545 550 36 696 DNA C. elegans coq-4 cDNA clk 36 atgtctgctc aaaagctata cgcatcgcat gttcccttgg caccgctgtc tcggatgctt 60 ctcgggattg gatcagcagt aacagcgatc tcggatccaa aaagaggaga tatggtggca 120 gcgatgggcg aaactactgc aattggacca gttttagaaa atattcgaaa aagaatggaa 180 tctgatgttg ttggaaagcg acttcttctc gaaaaaccaa gaatttcaaa tggaacaatt 240 gatagaaagt ggctaagaca gttaccggat ggaacattag gaaaattgta ttcaaacttt 300 ctcgatcgtt tgaacacatc tccagatgct cggcccactg tcaagtatat cgataatttg 360 gagcatcttt atgttatgca aaggtatcgc gaaacacacg acttcaccca catcgcattg 420 gagcagaaaa cgaacatgct cggcgaggta acagtcaaat atttcgaagg aattcaatat 480 gggcttccaa tgtgtgtcac tggtggaata tttggaggtg ccaggctttt aacaaaaaat 540 cgccaagaac ttgtcgaccg gaacctccct tgggttgtgg agcaggccac gaatgcacga 600 ttcttcatgg ctttcgactg ggaaaatcac tttgaaaagc agctcagcga ggtgcagaag 660 gagctaaata taacgccatt atctgtgaat atgtga 696 37 263 PRT unknown C. elegans COQ-4 protein 37 Met Ser Ala Gln Lys Leu Tyr Ala Ser His Val Pro Leu Ala Pro Leu 1 5 10 15 Ser Arg Met Leu Leu Gly Ile Gly Ser Ala Val Thr Ala Ile Ser Asp 20 25 30 Pro Lys Arg Gly Asp Met Val Ala Ala Met Gly Glu Thr Thr Ala Ile 35 40 45 Gly Pro Val Leu Glu Asn Ile Arg Lys Arg Met Glu Ser Asp Val Val 50 55 60 Gly Lys Arg Leu Leu Leu Glu Lys Pro Arg Ile Ser Asn Gly Thr Ile 65 70 75 80 Asp Arg Lys Trp Leu Arg Gln Leu Pro Asp Gly Thr Leu Gly Lys Leu 85 90 95 Tyr Ser Asn Phe Leu Asp Arg Leu Asn Thr Ser Pro Asp Ala Arg Pro 100 105 110 Thr Val Lys Tyr Ile Asp Asn Leu Glu His Leu Tyr Val Met Gln Arg 115 120 125 Tyr Arg Glu Thr His Asp Phe Thr His Ile Ala Leu Glu Gln Lys Thr 130 135 140 Asn Met Leu Gly Glu Val Thr Val Lys Tyr Phe Glu Gly Ile Gln Tyr 145 150 155 160 Gly Leu Pro Met Cys Val Thr Gly Gly Ile Phe Gly Gly Ala Arg Leu 165 170 175 Leu Thr Lys Asn Arg Gln Glu Leu Val Asp Arg Asn Leu Pro Trp Val 180 185 190 Val Glu Gln Ala Thr Asn Ala Arg Phe Phe Met Ala Phe Asp Trp Glu 195 200 205 Asn His Phe Glu Lys Gln Leu Ser Glu Val Gln Lys Glu Leu Asn Ile 210 215 220 Thr Pro Leu Ser Glu Leu Leu Asp Leu Pro Glu Met Glu Pro Asp Val 225 230 235 240 Pro Asp Ile Leu Phe Ser Lys Gly His Pro Gly Phe Trp Arg Val Leu 245 250 255 Gln Ala Val Asp Met Met Ile 260 38 268 PRT D. melanogaster COQ-4 38 Met Met Gln Arg Cys Leu Arg Leu Gln Lys Pro Leu Ala Leu Arg Arg 1 5 10 15 Gly Leu Arg Leu Ala Gln Ala Asn Ser Gln Ala Val Ala Thr Glu Ala 20 25 30 Pro Glu Ala Glu Pro Leu Asp Ala Phe Glu Arg Gln Tyr Leu Lys Glu 35 40 45 Arg Ile Glu Ile Ser Pro Phe Gln Arg Leu Phe Leu Gly Ala Gly Ser 50 55 60 Ser Ile Ala Ala Leu Leu Asn Pro Arg Arg His Asp Met Ile Ala Cys 65 70 75 80 Leu Gly Glu Thr Thr Gly Glu Asp Ala Leu Trp Thr Ile Leu Asp Thr 85 90 95 Met Gln Ala Ser Glu Glu Gly Gln Arg Ile Met Ala Asp Lys Pro Arg 100 105 110 Ile His Thr Ser Thr Ile Asp Phe Lys Tyr Leu Glu Thr Leu Pro Pro 115 120 125 Asp Thr Phe Gly Ala Ala Tyr Val Lys Phe Leu Lys Asp Asn Gln Val 130 135 140 Thr Pro Asp Ser Arg Met Ala Val Arg Phe Leu Glu Asp Pro Lys Leu 145 150 155 160 Ala Tyr Leu Met Thr Arg Tyr Arg Glu Cys His Asp Leu Ile His Thr 165 170 175 Val Leu Asp Met Pro Thr Asn Met Leu Gly Glu Val Ala Val Lys Trp 180 185 190 Val Glu Ala Leu Asn Thr Gly Leu Pro Met Cys Tyr Gly Gly Ala Val 195 200 205 Phe Gly Ala Val Arg Leu Arg Pro Lys Gln Arg Arg Ala Tyr Leu Lys 210 215 220 His Tyr Leu Pro Trp Ala Leu Glu Asn Gly Lys Arg Thr Lys Pro Leu 225 230 235 240 Met Pro Val Tyr Trp Glu Lys Arg Trp Glu Gln Asn Ile His Glu Leu 245 250 255 Arg Ser Glu Leu Gly Ile Thr Val Leu Asn Lys Ala 260 265 39 265 PRT Homo Sapiens COQ-4 39 Met Ala Thr Leu Leu Arg Pro Val Leu Arg Arg Leu Cys Gly Leu Pro 1 5 10 15 Gly Leu Gln Arg Pro Ala Ala Glu Met Pro Leu Arg Ala Arg Ser Asp 20 25 30 Gly Ala Gly Pro Leu Tyr Ser His His Leu Pro Thr Ser Pro Leu Gln 35 40 45 Lys Ala Leu Leu Ala Ala Gly Ser Ala Ala Met Ala Leu Tyr Asn Pro 50 55 60 Tyr Arg His Asp Met Val Ala Val Leu Gly Glu Thr Thr Gly His Arg 65 70 75 80 Thr Leu Lys Val Leu Arg Asp Gln Met Arg Arg Asp Pro Glu Gly Ala 85 90 95 Gln Ile Leu Gln Glu Arg Pro Arg Ile Ser Thr Ser Thr Leu Asp Leu 100 105 110 Gly Lys Leu Gln Ser Leu Pro Glu Gly Ser Leu Gly Arg Glu Tyr Leu 115 120 125 Arg Phe Leu Asp Val Asn Arg Val Ser Pro Asp Thr Arg Ala Pro Thr 130 135 140 Arg Phe Val Asp Asp Glu Glu Leu Ala Tyr Val Ile Gln Arg Tyr Arg 145 150 155 160 Glu Val His Asp Met Leu His Thr Leu Leu Gly Met Pro Thr Asn Ile 165 170 175 Leu Gly Glu Ile Val Val Lys Trp Phe Glu Ala Val Gln Thr Gly Leu 180 185 190 Pro Met Cys Ile Leu Gly Ala Phe Phe Gly Pro Ile Arg Leu Gly Ala 195 200 205 Gln Ser Leu Gln Val Leu Val Ser Glu Leu Ile Pro Trp Ala Val Gln 210 215 220 Asn Gly Arg Arg Ala His Cys Val Leu Asn Leu Tyr Tyr Glu Arg Arg 225 230 235 240 Trp Glu Gln Ser Leu Arg Ala Leu Arg Glu Glu Leu Gly Ile Thr Ala 245 250 255 Pro Pro Met His Val Gln Gly Leu Ala 260 265 40 272 PRT S. cerevisiae COQ-4 40 Met Phe Tyr Leu Asn Ala His Leu Glu Ile Asn Lys Val Val Asp Val 1 5 10 15 Val Met Ser Leu Ser Lys Lys Phe Leu Lys Pro Ser Val Ala Ser Asn 20 25 30 Gln Leu Arg Leu Leu Phe Thr Ala Ala Glu Arg Lys Val Asn Tyr Pro 35 40 45 Gly His Val Pro Leu Ser Pro Leu Gln Arg Ile Phe Leu Val Ala Gly 50 55 60 Ser Ala Ile Met Gly Leu Lys Ala Pro Trp Arg Gly Gly Asp Met Ile 65 70 75 80 Ser Val Leu Gly Asp Ala Ser Gly Gln Pro Phe Phe Leu His Arg Leu 85 90 95 Leu Asn Lys Met Leu Val Asp Lys Thr Gly Arg Glu Ile Leu Lys Asp 100 105 110 Lys Pro Arg Met Thr Ser Lys Ser Leu Asn Leu Pro Phe Leu Arg Thr 115 120 125 Leu Pro Pro Asn Thr Leu Gly Lys Ile Tyr Val Asp Trp Ile Asp Lys 130 135 140 Glu His Val Gly Pro Asp Thr Arg Ser Pro Thr Arg Phe Val Asp Asp 145 150 155 160 Pro Glu Glu Ala Tyr Val Met Gln Arg Tyr Arg Glu Ser His Asp Phe 165 170 175 Tyr His Ala Ile Cys Asn Met Pro Thr Asn Ile Glu Gly Glu Leu Ala 180 185 190 Ile Lys Trp Leu Glu Phe Val Asn Met Gly Leu Pro Val Gly Ala Leu 195 200 205 Ser Ala Leu Phe Gly Pro Leu Arg Leu Asn Cys Glu Gln Ala Ser Arg 210 215 220 Phe Arg Arg Val Tyr Ile Pro Trp Ser Ile Arg Asn Gly Leu Asn Ala 225 230 235 240 Lys Thr Leu Ile Asn Val Tyr Trp Glu Lys Glu Leu Thr Asn Asp Ile 245 250 255 Glu Asp Val Arg Arg Arg Ile Arg Ile Glu Ala Ala Pro Pro Leu Val 260 265 270 41 226 PRT Arabidopsis thaliana COQ-4 41 Met Ile Ile Glu Arg Ala Arg Val Pro Leu Ser Arg Trp Gln Gln Ala 1 5 10 15 Ala Val Ala Met Gly Ser Ala Leu Gly Ala Leu Val Asp Pro Arg Arg 20 25 30 Ala Asp Leu Ile Ala Ala Leu Gly Glu Thr Thr Gly Lys Pro Ala Phe 35 40 45 Glu Met Val Leu Glu Arg Met Lys Lys Ser Glu Glu Gly Arg Ala Ile 50 55 60 Leu Leu Glu Arg Pro Arg Val Val Ser Glu Gln Val Gly His Ala Trp 65 70 75 80 Asp Leu Pro Glu Asn Thr Phe Gly Ala Ala Tyr Ala Lys Phe Met Gly 85 90 95 Ser Arg Asn Phe Ser Pro Asp Asp Arg Pro Pro Val Arg Phe Met Glu 100 105 110 Thr Asp Glu Leu Ala Tyr Val Ala Thr Arg Ala Arg Glu Val His Asp 115 120 125 Leu Trp His Thr Leu Phe Gly Leu Pro Thr Asn Leu Ile Gly Glu Ser 130 135 140 Ser Leu Lys Val Ile Glu Phe Glu Gln Met Tyr Leu Pro Met Cys Met 145 150 155 160 Leu Ser Val Ile Gly Gly Thr Val Arg Phe Asn Glu Lys Gln Arg Ser 165 170 175 Met Phe Leu Lys His Tyr Leu Pro Trp Ala Val Arg Ala Gly Arg Gln 180 185 190 Cys Thr Asp Leu Met Cys Val Tyr Tyr Glu Arg His Phe Ser Glu Asp 195 200 205 Leu Glu Gln Val Arg Arg Lys Trp Gly Ile Ile Pro Ala Pro Gln His 210 215 220 Pro Lys 225 42 167 PRT unknown part of Mus musculus COQ-4 protein 42 Ala Leu Tyr Asn Pro Tyr Arg His Asp Met Val Ala Val Leu Gly Glu 1 5 10 15 Thr Thr Gly Cys His Thr Leu Lys Phe Leu Arg Asp Gln Met Lys Lys 20 25 30 Asp Pro Glu Gly Ala Gln Ile Leu Gln Glu Arg Pro Arg Ile Ser Leu 35 40 45 Ser Thr Leu Asp Leu Ser Lys Leu Gln Ser Leu Pro Glu Gly Ser Leu 50 55 60 Gly Arg Glu Tyr Leu Arg Phe Leu Asp Val Asn Lys Val Ser Pro Asp 65 70 75 80 Thr Arg Ala Pro Thr Arg Phe Val Asp Asp Glu Glu Leu Ala Tyr Val 85 90 95 Ile Gln Arg Tyr Arg Glu Val His Asp Met Leu His Thr Leu Leu Gly 100 105 110 Met Pro Thr Asn Met Leu Gly Glu Val Val Val Lys Trp Phe Glu Ala 115 120 125 Val Gln Thr Gly Leu Pro Met Cys Ile Leu Gly Ala Leu Phe Gly Pro 130 135 140 Ile Arg Leu Arg Thr Gln Ser Leu Gln Val Leu Phe Ser Glu Leu Ile 145 150 155 160 Pro Trp Ala Ile Gln Asn Gly 165 43 123 PRT unknown part of Mus musculus COQ-4 protein 43 Tyr Pro Asp His Ile Pro Thr Thr Pro Leu Gln Lys Met Leu Leu Ala 1 5 10 15 Ala Gly Ala Ala Gly Met Ala Leu Tyr Asn Pro Tyr Arg His Asp Met 20 25 30 Val Ala Val Leu Gly Glu Thr Thr Gly Cys His Thr Leu Lys Phe Leu 35 40 45 Arg Asp Gln Met Lys Lys Asp Pro Glu Gly Ala Gln Ile Leu Gln Glu 50 55 60 Arg Pro Arg Ile Ser Leu Ser Thr Leu Asp Leu Ser Lys Leu Gln Ser 65 70 75 80 Leu Pro Glu Gly Ser Leu Gly Arg Glu Tyr Leu Arg Phe Leu Asp Val 85 90 95 Asn Lys Val Ser Pro Asp Thr Arg Ala Pro Thr Arg Phe Val Asp Asp 100 105 110 Glu Xaa Leu Ala Tyr Val Asn Gln Lys Tyr Arg 115 120 44 89 PRT unknown part of Mus musculus COQ-4 protein 44 His Leu Pro Thr Ser Tyr Pro Ser Leu Ser Leu Gly Glu Val Val Val 1 5 10 15 Lys Trp Phe Glu Ala Val Gln Thr Gly Leu Pro Met Cys Ile Leu Gly 20 25 30 Ala Leu Phe Gly Pro Ile Arg Leu Arg Thr Gln Ser Leu Gln Val Leu 35 40 45 Phe Ser Glu Leu Ile Pro Trp Ala Ile Gln Asn Gly Arg Arg Ala Thr 50 55 60 Cys Val Leu Asn Ile Tyr Tyr Glu Gln Arg Trp Glu Gln Pro Leu Thr 65 70 75 80 Ala Leu Arg Glu Glu Leu Gly Ile Ser 85 45 218 PRT Unknown Mus musculs consensus COQ-4 proteein 45 Tyr Pro Asp His Ile Pro Thr Thr Pro Leu Gln Lys Met Leu Leu Ala 1 5 10 15 Ala Gly Ala Ala Gly Met Ala Leu Tyr Asn Pro Tyr Arg His Asp Met 20 25 30 Val Ala Val Leu Gly Glu Thr Thr Gly Cys His Thr Leu Lys Phe Leu 35 40 45 Arg Asp Gln Met Lys Lys Asp Pro Glu Gly Ala Gln Ile Leu Gln Glu 50 55 60 Arg Pro Arg Ile Ser Leu Ser Thr Leu Asp Leu Ser Lys Leu Gln Ser 65 70 75 80 Leu Pro Glu Gly Ser Leu Gly Arg Glu Tyr Leu Arg Phe Leu Asp Val 85 90 95 Asn Lys Val Ser Pro Asp Thr Arg Ala Pro Thr Arg Phe Val Asp Asp 100 105 110 Glu Glu Leu Ala Tyr Val Ile Gln Arg Tyr Arg Glu Val His Asp Met 115 120 125 Leu His Thr Leu Leu Gly Met Pro Thr Asn Met Leu Gly Glu Val Val 130 135 140 Val Lys Trp Phe Glu Ala Val Gln Thr Gly Leu Pro Met Cys Ile Leu 145 150 155 160 Gly Ala Leu Phe Gly Pro Ile Arg Leu Arg Thr Gln Ser Leu Gln Val 165 170 175 Leu Phe Ser Glu Leu Ile Pro Trp Ala Ile Gln Asn Gly Arg Arg Ala 180 185 190 Thr Cys Val Leu Asn Ile Tyr Tyr Glu Gln Arg Trp Glu Gln Pro Leu 195 200 205 Thr Ala Leu Arg Glu Glu Leu Gly Ile Ser 210 215 46 137 PRT Glycine max COQ-4 46 Leu Pro Ala Asn Thr Phe Gly Ala Ala Tyr Ala Arg Phe Met Gly Ser 1 5 10 15 Arg Asn Phe Ser Pro Asp Asp Arg Pro Pro Val Arg Phe Met Asp Thr 20 25 30 Asp Glu Leu Ala Tyr Val Ala Met Arg Ala Arg Glu Val His Asp Phe 35 40 45 Trp His Thr Leu Phe Asp Leu Pro Thr Asn Leu Ile Gly Glu Thr Ala 50 55 60 Leu Lys Val Ile Glu Phe Glu Gln Met Gly Leu Pro Met Cys Leu Leu 65 70 75 80 Ser Val Ile Gly Gly Thr Ala Arg Phe Ser Glu Lys Gln Arg Lys Leu 85 90 95 Phe Tyr His His Tyr Phe Pro Trp Ala Ile His Ala Gly Met Pro Ser 100 105 110 Thr Asp Leu Met Cys Val Tyr Tyr Glu Arg His Phe Asp Glu Asp Leu 115 120 125 Glu Asp Val Arg Arg Lys Leu Gln Ile 130 135 47 136 PRT Bos taurus COQ-4 47 Tyr Pro Glu His Ile Pro Thr Ser Val Leu Gln Lys Val Leu Leu Ala 1 5 10 15 Ala Gly Ser Ala Gly Met Ala Leu Tyr Asp Pro Tyr Arg His Asp Met 20 25 30 Val Ala Val Leu Gly Glu Thr Thr Gly Arg Arg Thr Leu Lys Val Leu 35 40 45 Arg Asp Gln Met Lys Arg Asp Pro Glu Gly Ala Gln Ile Leu Gln Glu 50 55 60 Arg Pro Arg Ile Ser Leu Ser Thr Leu Asp Met Gly Lys Leu Arg Ser 65 70 75 80 Leu Pro Glu Gly Ser Phe Gly Cys Ala Tyr Leu His Phe Leu Asp Val 85 90 95 Asn Arg Val Ser Pro Asp Thr Arg Ala Pro Thr Arg Phe Val Asp Asp 100 105 110 Glu Glu Leu Ala Tyr Val Ile Gln Arg Tyr Arg Glu Ile His Asp Met 115 120 125 Leu His Ala Leu Leu Gly Met Pro 130 135 48 120 PRT Medicago trunculata COQ-4 48 Asn Phe Ser Pro Asp Asp Arg Pro Pro Val Arg Phe Met Asp Thr Asp 1 5 10 15 Glu Leu Ala Tyr Val Ala Met Arg Ala Arg Glu Val His Asp Phe Trp 20 25 30 His Thr Leu Phe Asp Leu Pro Thr Asn Leu Ile Gly Glu Ser Ala Leu 35 40 45 Lys Val Ile Glu Phe Glu Gln Met His Leu Pro Met Cys Val Met Ser 50 55 60 Val Leu Gly Gly Thr Ala Arg Phe Ser Glu Lys Gln Arg Lys Leu Phe 65 70 75 80 Tyr Gln His Tyr Phe Pro Trp Ala Val Arg Ala Gly Thr Gln Cys Asn 85 90 95 Asp Leu Met Cys Val Tyr Tyr Glu Gln His Phe His Gln Asp Leu Glu 100 105 110 Asp Val Arg Arg Lys Leu Gly Ile 115 120 49 113 PRT Ancylostoma caninum COQ-4 49 Lys Phe Val Gln Asn Ser Glu His Leu Tyr Val Met Gln Arg Tyr Arg 1 5 10 15 Glu Thr His Asp Phe Asn His Val Leu Leu Gln Met Pro Thr His Met 20 25 30 Leu Gly Glu Val Thr Val Lys Tyr Phe Glu Gly Ile Gln Phe Gly Leu 35 40 45 Pro Met Cys Val Thr Ala Gly Leu Phe Gly Ala Ala Arg Leu Arg Lys 50 55 60 Asn His Arg His Arg Phe Leu Thr Gln His Leu Pro Trp Ile Val Glu 65 70 75 80 Gln Ala Thr Lys Gly Arg Phe Phe Met Ala Ile Asp Trp Glu Asn His 85 90 95 Trp Glu Glu Thr Ile Pro Ser Leu Gln Glu Gln Phe Gly Ile Thr Pro 100 105 110 Leu 50 113 PRT Trypanosoma cruzi COQ-4 VARIANT (1)...(113) Xaa = Any Amino Acid 50 Phe Val Ala Ala Thr Thr Arg Ser Ile Trp Asp Pro Val Asn Ala Asp 1 5 10 15 Asp Val Ala Ala Val Gly Glu Thr Pro Ala Leu Thr Ala Leu Gly His 20 25 30 Met Lys Gln Ser Met Met Ser Asp Arg Thr Gly Arg Met Ile Leu Arg 35 40 45 Thr Gln Pro Arg Val Thr Asp Glu Thr Leu Glu Phe Ala Ser Arg Gln 50 55 60 Pro Pro Gly Thr Phe Gly His Arg Tyr Ala Gln Phe Met Lys Phe Xaa 65 70 75 80 Xaa Phe Thr Pro Asn Gly Arg Thr Pro Val Ala His Ile Ala Asp Pro 85 90 95 Thr Leu Ala Tyr Val Met Gln Arg Gln Arg Glu Thr His Asp Phe Leu 100 105 110 His 51 99 PRT Rattus rattus COQ-4 51 His Pro Tyr Arg His Asp Met Leu Pro Val Leu Gly Glu Thr Thr Gly 1 5 10 15 Cys His Thr Leu Lys Phe Leu Arg Asp Gln Met Lys Lys Asp Pro Glu 20 25 30 Gly Ala Gln Ile Leu Gln Glu Arg Pro Arg Ile Ser Leu Ser Thr Leu 35 40 45 Asp Leu Ser Lys Leu Gln Ser Leu Pro Glu Gly Ser Leu Gly Arg Glu 50 55 60 Tyr Leu Arg Phe Leu Asn Ala Asn Lys Val Ser Pro Asp Thr Arg Ala 65 70 75 80 Pro Thr Arg Phe Val Asp Asp Glu Glu Leu Ala Tyr Val Ile Gln Arg 85 90 95 Tyr Arg Glu 52 122 PRT Gossypium hirsutum COQ-4 VARIANT (1)...(122) Xaa = Any Amino Acid 52 Ile Lys Leu Ser Pro Trp Gln Gln Ala Ala Val Ala Val Gly Ser Ala 1 5 10 15 Val Gly Ala Leu Leu Asp Pro Arg Arg Ala Asp Leu Ile Ala Ala Leu 20 25 30 Gly Glu Thr Thr Gly Lys Pro Ala Phe Glu Arg Val Leu Glu Arg Met 35 40 45 Arg Arg Ser Pro Glu Gly Lys Thr Xaa Leu Leu Glu Arg Pro Arg Val 50 55 60 Ile Ser Ala Asn Val Gly His Ala Trp Asp Leu Pro Lys Asn Thr Phe 65 70 75 80 Gly Ala Ala Tyr Ala Arg Phe Leu Gly Ser Xaa Asn Phe Ser Pro Asp 85 90 95 Asp Arg Pro Pro Val Arg Phe Met Asp Thr Asp Glu Leu Ala Tyr Val 100 105 110 Ala Met Arg Ala Arg Glu Val His Asp Phe 115 120 

What is claimed is:
 1. A clk-2 gene which has a function at the level of cellular physiology involved in developmental rate, telomere length and longevity, wherein clk-2 mutations cause a longer life, an altered cellular metabolism and/or an altered telomere length relative to the wild type, wherein clk-2 overexpression leads to telomere shortening, and wherein clk-2 gene has the identifying characteristics of nucleotide sequence set forth in SEQ ID NO:1.
 2. Use of a clk-2 gene to alter a function at the level of cellular physiology involved in the regulation of developmental rates, telomere length and longevity, wherein clk-2 mutations cause a longer life, altered cellular metabolism and physiological rates and/or an altered telomere length relative to the wild type, wherein clk-2 overexpression leads to telomere shortening, and wherein clk-2 gene has the identifying characteristics of nucleotide sequences set forth in SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, or SEQ ID NO:24, or wherein said gene codes for a protein sequence as set forth in SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31 or SEQ ID NO:32.
 3. The clk-2 gene of claim 1, which codes for a CLK-2 protein having the amino acid sequence set forth in SEQ ID NO:2.
 4. The use of a clk-2 gene to alter function at the level of cellular physiology involved in the regulation of developmental rates, telomere length and/or longevity, wherein clk-2 mutations cause a longer life, altered cellular metabolism and physiological rates and an altered telomere length relative to the wild type, wherein clk-2 overexpression leads to telomere shortening, and wherein said gene codes for a protein having a sequence as set forth in SEQ ID NO:32.
 5. A CLK-2 protein which has a function at the level of cellular physiology involved in the regulation of developmental rate, telomere length and longevity, wherein said CLK-2 protein is encoded by the gene of claim
 1. 6. Use of a CLK-2 protein to alter a function at the level of cellular physiology involved in the regulation of developmental rate, telomere length and longevity, wherein clk-2 overexpression leads to telomere shortening, and wherein said CLK-2 protein is encoded by a gene as defined in claim
 2. 7. A mutant CLK-2 protein which has the amino acid sequence set forth in SEQ ID NO:31.
 8. A CLK-2 protein which has the amino acid sequence set forth in SEQ ID NO:2.
 9. Use of CLK-2 protein to alter a function at the level of cellular physiology involved in the regulation of developmental rates, telomere length and longevity, wherein said CLK-2 protein has the amino acid sequence as set forth in SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31 or SEQ ID NO:32.
 10. A clk-2 gene which has the nucleotide sequence set forth in SEQ ID NO:1.
 11. A mouse which comprises a gene knockout of the murine clk-2 gene homologue to a clk-2 gene as defined in claim
 2. 12. A method to increase the life span of multicellular organism which comprises altering the function of telomeres and/or regulating telomere length.
 13. The method of claim 12, wherein said multicellular organism is a metazoan.
 14. A method to increase the life span of multicellular organism which comprises altering the mechanisms of sub-telomeric silencing and/or regulating telomere length.
 15. The method of claim 14, wherein said multicellular organism is a metazoan.
 16. The use of clk-2 gene as defined in claim 1, 3 or 10 and homologues thereof, to manipulate the physiological rates and/or telomere biology, whereby life span of an organism is altered.
 17. Use of elk-2 gene as defined in claim 1, 3 or 10, or CLK-2 protein as defined in claim 5, 7 or 8 and homologues thereof, for screening drugs which decrease or increase the life span of a multicellular organism.
 18. The use of claim 17, wherein said drug enhances or suppresses the expression of the clk-2 gene or activity of the protein CLK-2, and homologues thereof.
 19. Use of a compound for the manufacture of a medicament for increasing and/or decreasing physiological rates of tissues, organ, and/or whole organism of a host; wherein said compound is interfering with activity of CLK-2 protein of claim 5, 7 or 8, and homologues thereof.
 20. Use of a compound which promotes tissue and/or organ specific reduction or increase of clk-2 activity for the manufacture of a medicament for the treatment of pathological conditions causing increase of physiological rate of tissue and/or organ in an individual, wherein said compound is interfering with activity of CLK-2 protein of claim 5, 7 or 8, and homologues thereof.
 21. Use of a compound which promotes tissue and/or organ specific reduction or increase of clk-2 activity for the manufacture of a medicament for the treatment of pathological conditions causing decrease of physiological rate of tissue and/or organ in an individual, wherein said compound is interfering with activity of CLK-2 protein as defined in claim 5, 7 or 8, and homologues thereof.
 22. A clk-2 co-expressed gene which comprises a cex-7 gene having the nucleotide sequence set forth in SEQ ID NO:33 which codes for a CEX-7 protein having the amino acid sequence set forth in SEQ ID NO:34 wherein said gene is located in the clk-2 operon and said cex-7 gene is transcriptionally co-expressed with clk-2 gene present in said operon.
 23. A human homologue of cex-7 gene of claim 22, wherein said gene codes for a protein having a sequence as set forth in SEQ ID NO:35.
 24. Use of a human homologue of cex-7 gene of claim 22 and homologues thereof, to alter a function at the level of cellular level physiology involved in the regulation of developmental rates and longevity wherein said gene codes for a protein having a sequence as set forth in SEQ ID NO:35.
 25. A mouse which comprises a gene knock out of the murine cex-7 gene homologue of the human gene as set forth in SEQ ID NO:35.
 26. Use of a compound for the manufacture of a medicament for increasing and/or decreasing physiological rates of tissues, organ, and/or whole organism of a host; wherein said compound is interfering with activity of CEX-7 as defined in claim 22 or 23, and homologues thereof.
 27. Use of a compound which promotes tissue and/or organ specific reduction or increase of cex-7 activity for the manufacture of a medicament for the treatment of pathological conditions causing increase of physiological rate of tissue and/or organ in an individual, wherein said compound is interfering with activity of CEX-7 as defined in claim 22 or 23, and homologues thereof.
 28. Use of a compound which promotes tissue and/or organ specific reduction or increase of cex-7 activity for the manufacture of a medicament for the treatment of pathological conditions causing decrease of physiological rate of tissue and/or organ in an individual, wherein said compound is interfering with activity of CEX-7 as defined in claim 22 or 23, and homologues thereof.
 29. A coq-4 gene which has a function at the level of cellular physiology involved in the regulation of. developmental rate and longevity, wherein coq-4 mutations cause altered cellular metabolism and physiological relative to the wild type, wherein coq-4 gene has the identifying characteristics of nucleotide sequence set forth in SEQ ID NO:36.
 30. A coq-4 gene which has a function at the level of cellular physiology involved in the regulation of developmental rate and longevity, wherein coq-4 mutations cause altered cellular metabolism and physiological relative to the wild type, wherein coq-4 gene has the identifying characteristics of nucleotide sequence set forth in SEQ ID NO:36, wherein said gene codes for a protein having a sequence as set forth in SEQ ID NO:37.
 31. Use of coq-4 gene to alter a function at the level of cellular physiology involved in the regulation of developmental rates, wherein coq-4 mutations cause an altered cellular metabolism and physiological rates relative to the wild type, wherein said gene codes for a protein having a sequence as set forth in SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51 or SEQ ID NO:52 and homologues thereof.
 32. A COQ-4 protein which has a function at the level of cellular physiology involved in the regulation of developmental rate and longevity, wherein said COQ-4 protein is encoded by the gene of claim
 29. 33. A mouse which comprises a gene knock out of the murine coq-4 gene as set forth in SEQ ID NO:45.
 34. Use of a compound for the manufacture of a medicament for increasing and/or decreasing physiological rates of tissues, organs and/or whole organism of a host; wherein said compound is interfering with activity of COQ-4 protein as defined in claim 32, and homologues thereof.
 35. Use of a compound which promotes tissue and/or organ specific reduction or increase of coq-4 activity for the manufacture of a medicament for the treatment of pathological conditions causing increase of physiological rate of tissue and/or organ in an individual, wherein said compound is interfering with activity of COQ-4 protein as defined in claim 32, and homologues thereof.
 36. Use of a compound which promotes tissue and/or organ specific reduction or increase of coq-4 activity for the manufacture of a medicament for the treatment of pathological conditions causing decrease of physiological rate of tissue and/or organ in an individual, wherein said compound is interfering with activity of COQ-4 protein as defined in claim 32, and homologues thereof.
 37. Use of a compound which promotes tissue and/or organ specific reduction or increase of clk-2 activity for the manufacture of a medicament for the treatment of pathological conditions due to altered telomere length in tissue and/or organ in an individual, wherein said compound is interfering with activity of CLK-2 protein as defined in claim 5, 7 or 8, and homologues thereof. 